Synthetic cytokine receptors

Abstract

Synthetic cytokine receptors are provided herein, each comprising an extracellular domain, a transmembrane domain, and an interleukin-9 receptor (IL-9R) intracellular domain. Modified cells expressing one or more such synthetic cytokine receptors are also provided herein.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims priority based on U.S. Provisional Patent Application No. 63 / 510,921, filed on June 29, 2023, entitled "SYNTHETIC CYTOKINE RECEPTORS"; U.S. Provisional Patent Application No. 63 / 615,248, filed on December 27, 2023, entitled "Synthetic Cytokine Receptors"; and U.S. Provisional Patent Application No. 63 / 555,875, filed on February 20, 2024, entitled "Synthetic Cytokine Receptors", the contents of which are hereby incorporated by reference in their entirety.

[0002] Reference to Electronic Sequence Listing This application is filed with an electronic sequence listing. The sequence listing is provided as a file named 30761 - 2000240.XML, created on June 28, 2024, with a size of 277,183 bytes. The electronic form information of the sequence listing is hereby incorporated by reference in its entirety.

[0003] Field This disclosure generally relates to synthetic cytokine receptors and modified cells that express one or more such synthetic cytokine receptors.

Background Art

[0004] Background Therapies using adoptive transfer of genetically modified T cells have shown significant antitumor activity in patients with hematologic malignancies. However, such therapies have limited benefit in patients with solid tumors. One major limitation is the insufficient in vivo expansion and persistence of the adoptively transferred T cells. To circumvent this limitation, patients receive lymphodepletion by chemotherapy and / or radiation prior to the transfer of modified T cells. However, there are also patients who are too debilitated to receive toxic regimens such as chemotherapy and radiation. Another major limitation is that T cells that expand and persist successfully in vivo become terminally differentiated and dysfunctional. Thus, improved systems and modified cells are needed to address these problems. The provided embodiments address these needs. SUMMARY OF THE INVENTION

[0005] SUMMARY The present disclosure provides synthetic cytokine receptors, modified cells that express one or more such synthetic cytokine receptors, and uses thereof.

[0006] In some embodiments, synthetic cytokine receptors are provided herein that comprise an extracellular domain, a transmembrane domain, and an interleukin 9 receptor (IL-9R) intracellular domain.

[0007] In some embodiments, polynucleotides encoding synthetic cytokine receptors that comprise an extracellular domain, a transmembrane domain, and an interleukin 9 receptor (IL-9R) intracellular domain are provided herein.

[0008] In some embodiments, vectors are provided herein that comprise polynucleotides encoding synthetic cytokine receptors that comprise an extracellular domain, a transmembrane domain, and an interleukin 9 receptor (IL-9R) intracellular domain.

[0009] In some embodiments, methods for modifying isolated cells are provided herein, comprising the step of contacting cells with a polynucleotide encoding a synthetic cytokine receptor comprising an extracellular domain, a transmembrane domain, and an interleukin-9 receptor (IL-9R) intracellular domain, or a vector comprising such a polynucleotide.

[0010] In some embodiments, modified cells expressing one or more synthetic cytokine receptors comprising an extracellular domain, a transmembrane domain, and an interleukin-9 receptor (IL-9R) intracellular domain are provided herein.

[0011] In some embodiments, a cell population is provided herein that includes at least one modified cell expressing one or more synthetic cytokine receptors comprising an extracellular domain, a transmembrane domain, and an interleukin-9 receptor (IL-9R) intracellular domain.

[0012] In some embodiments, the following pharmaceutical compositions are provided herein, comprising modified cells expressing one or more synthetic cytokine receptors comprising an extracellular domain, a transmembrane domain, and an interleukin-9 receptor (IL-9R) intracellular domain, or a population of cells comprising at least one such modified cell.

[0013] In some embodiments, methods for treating cancer in a subject are provided herein, comprising the step of administering a therapeutically effective amount of a pharmaceutical composition disclosed herein to the subject.

[0014] In some aspects, synthetic cytokine receptors are provided herein that are homodimers of the same polypeptide chain, each containing an extracellular domain, a transmembrane domain, and an intracellular domain capable of interleukin-9 receptor (IL-9R) signaling.

[0015] In some embodiments, the IL-9R signaling-capable intracellular domain comprises an IL-9R intracellular domain or a variant thereof. In some embodiments, the IL-9R signaling-capable intracellular domain comprises a chimeric JAK / STAT fusion domain.

[0016] In some aspects, synthetic cytokine receptors are provided herein that are constitutively active cytokine receptors comprising an extracellular domain, a transmembrane domain, and an interleukin-9 receptor (IL-9R) intracellular domain or a variant thereof.

[0017] In some embodiments, the synthetic cytokine receptor is a multimer. In some embodiments, the synthetic cytokine receptor is a multimer of identical polypeptide chains, each containing an extracellular domain, a transmembrane domain, and an IL-9R intracellular domain or a variant thereof. In some embodiments, the multimer is a dimer. In some embodiments, the dimer is a homodimer. In some embodiments, each polypeptide chain is constitutively multimerized.

[0018] In some embodiments, the synthetic cytokine receptor comprises at least one self-assembling domain. In some embodiments, the at least one self-assembling domain is an extracellular domain and / or a transmembrane domain.

[0019] In some embodiments, synthetic cytokine receptors are multimerized via transmembrane and / or extracellular domains.

[0020] In some aspects, synthetic cytokine receptors are provided herein that are homodimers of the same polypeptide chain, each containing an extracellular domain, a transmembrane domain, and an intracellular domain of the interleukin-9 receptor (IL-9R) or a variant thereof.

[0021] In some embodiments, the extracellular domain and / or transmembrane domain are heterogeneous to IL-9R. In some embodiments, the transmembrane domain and extracellular domain are derived from the same protein. In some embodiments, the transmembrane domain and extracellular domain are derived from different proteins.

[0022] In some embodiments, the transmembrane domain is 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 amino acids long. In some embodiments, the transmembrane domain includes a transmembrane domain derived from glycophorin A (GpA), carnitine palmitoyltransferase 1 (CPT1), tumor necrosis factor receptor (TNFR), or Muc24. In some embodiments, the transmembrane domain includes a transmembrane domain derived from the thrombopoietin receptor.

[0023] In some embodiments, the transmembrane domain promotes α-helix dimerization. In some embodiments, the transmembrane domain contains motif GXXXG (SEQ ID NO:68). In some embodiments, the transmembrane domain is a motif Includes TIFF2026525229000001.tif4128.

[0024] In some embodiments, the transmembrane domain is a transmembrane domain derived from glycophorin A (GpA), or a variant thereof containing one or more mutations (e.g., 1, 2, 3, 4, 5, or 6 mutations) compared to the wild-type GpA transmembrane domain, wherein the variant GpA is sufficient to promote α-helix dimerization. In some embodiments, the transmembrane domain comprises a transmembrane domain derived from glycophorin A (GpA). In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:22 or SEQ ID NO:43. In some embodiments, the transmembrane domain contains the amino acid sequence of SEQ ID NO:22 or SEQ ID NO:43.

[0025] In some embodiments, the transmembrane domain includes a GXXXG (SEQ ID NO:68) motif and a GXXXA (SEQ ID NO:69) motif. In some embodiments, the transmembrane domain includes a transmembrane domain derived from carnitine palmitoyltransferase 1 (CPT1). In some embodiments, the transmembrane domain includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:45. In some embodiments, the transmembrane domain includes the amino acid sequence of SEQ ID NO:45.

[0026] In some embodiments, the synthetic cytokine receptor comprises motif AXXXA (SEQ ID NO: 77) or AXXXS (SEQ ID NO: 78). In some embodiments, the synthetic cytokine receptor comprises motif ΦPXΦ (SEQ ID NO: 75) or ΦTXXAΦ (SEQ ID NO: 76). In some embodiments, the transmembrane domain is derived from TNFR. In some embodiments, TNFR is TACI, DR5, p75NTR, Fas, TNFR1, TNFR2, or OX40.

[0027] In some embodiments, the transmembrane domain includes a transmembrane domain derived from DR5. In some embodiments, the transmembrane domain includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:46. In some embodiments, the transmembrane domain includes the amino acid sequence of SEQ ID NO:46.

[0028] In some embodiments, the transmembrane domain includes a transmembrane domain derived from TACI. In some embodiments, the transmembrane domain includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:44. In some embodiments, the transmembrane domain includes the amino acid sequence of SEQ ID NO:44.

[0029] In some embodiments, the transmembrane domain contains 1 to 6 cysteine ​​residues. In some embodiments, the transmembrane domain promotes disulfide dimerization. In some embodiments, disulfide dimerization forms 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

[0030] In some embodiments, the transmembrane domain is a variant transmembrane domain containing one or more mutations compared to the wild-type transmembrane domain to promote homodimerization of the receptor polypeptide.

[0031] In some embodiments, one or more mutations promote α-helix dimerization or disulfide bond dimerization. In some embodiments, one or more mutations introduce at least one cysteine ​​into the transmembrane domain. In some embodiments, one or more mutations introduce proline into the transmembrane domain. In some embodiments, one or more mutations introduce threonine into the transmembrane domain.

[0032] In some embodiments, one or more mutations introduce a trimer peptide of cysteine, proline, and another amino acid other than cysteine ​​and proline into the transmembrane domain. In some embodiments, one or more mutations introduce a trimer peptide of cysteine, proline, and threonine (CPT or TCP) into the transmembrane domain.

[0033] In some embodiments, the transmembrane domain is a variant IL-7R transmembrane domain, and one or more mutations are in the wild-type transmembrane sequence. It is located in TIFF2026525229000002.tif4128. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:21. In some embodiments, the transmembrane domain contains the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:21.

[0034] In some embodiments, the transmembrane domain includes a transmembrane domain derived from Muc24. In some embodiments, the transmembrane domain includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:23. In some embodiments, the transmembrane domain includes the amino acid sequence of SEQ ID NO:23.

[0035] In some embodiments, the extracellular domain is approximately 150–260 amino acids long.

[0036] In some embodiments, the extracellular domain is a dimerization domain. In some embodiments, the dimerization domain includes a hinge region. In some embodiments, the extracellular domain promotes disulfide bond dimerization. In some embodiments, the extracellular domain contains 1 to 6 cysteine ​​residues. In some embodiments, disulfide bond dimerization forms 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

[0037] In some embodiments, the extracellular domain is derived from the extracellular domain of CD34, DAP12, glycophorin A, CD8, or Muc24. In some embodiments, the extracellular domain includes an extracellular domain derived from the thrombopoietin receptor.

[0038] In some embodiments, the extracellular domain comprises the extracellular domain of CD8, or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain comprises an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:18 or SEQ ID NO:19. In some embodiments, the extracellular domain comprises the amino acid sequence of SEQ ID NO:18 or SEQ ID NO:19.

[0039] In some embodiments, the extracellular domain comprises the extracellular domain of CD34, or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain comprises an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:4. In some embodiments, the extracellular domain comprises the amino acid sequence of SEQ ID NO:4.

[0040] In some embodiments, the extracellular domain is the extracellular domain of Muc24, or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:20. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:20.

[0041] In some embodiments, the extracellular domain is the extracellular domain of DAP12, or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:5. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:5.

[0042] In some embodiments, the extracellular domain is the extracellular domain of glycophorin A (GpA), or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:16 or SEQ ID NO:17. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:16 or SEQ ID NO:17.

[0043] In some embodiments, the IL-9R intracellular domain or its variant is approximately 100 to 260 amino acids long. In some embodiments, the IL-9R intracellular domain or its variant contains a box 1 motif and / or a box 2 motif. In some embodiments, the IL-9R intracellular domain or its variant contains a box 2 motif. In some embodiments, the IL-9R intracellular domain or its variant is 230 amino acids long.

[0044] In some embodiments, the IL-9R intracellular domain is the wild-type IL-9R intracellular domain, or a variant thereof containing one or more mutations compared to the wild-type IL-9R intracellular domain shown in SEQ ID NO:8. In some embodiments, the one or more mutations include one or more amino acid insertions, deletions, and / or substitutions. In some embodiments, the one or more mutations promote signaling through the STAT1, STAT3, and / or STAT5 pathways. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence of SEQ ID NO:8.

[0045] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, or SEQ ID NO:56.

[0046] In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:67. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence of SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:67.

[0047] In some embodiments, the IL-9R intracellular domain or its variants contain one or more amino acid deletions relative to the wild-type IL-9R intracellular domain (SEQ ID NO:8).

[0048] In some embodiments, the IL-9R intracellular domain or its variant is a shortened IL-9R lacking the continuous amino acid sequence at the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the shortened IL-9R intracellular domain or its variant is shortened by only 62 to 99 consecutive amino acids from the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the IL-9R intracellular domain or its variant is a shortened IL-9R lacking amino acids 132 to 230 of SEQ ID NO:8 or amino acids 134 to 230 of SEQ ID NO:8.

[0049] In some embodiments, the IL-9R intracellular domain or its variants contain an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:54, SEQ ID NO:103, or SEQ ID NO:104.

[0050] In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, or SEQ ID NO:63.

[0051] In some embodiments, the IL-9R intracellular domain or its variants contain one or more amino acid substitutions relative to the wild-type IL-9R intracellular domain (SEQ ID NO:8).

[0052] In some embodiments, the IL-9R intracellular domain or a variant thereof includes a STAT binding motif or a variant thereof. In some embodiments, the STAT binding motif includes a STAT1 binding motif, a STAT3 binding motif, and / or a STAT5 binding motif. In some embodiments, the STAT binding motif includes YLPQ (SEQ ID NO: 171). In some embodiments, the STAT binding motif includes a variant STAT binding motif. In some embodiments, the variant STAT binding motif includes YRPQ (SEQ ID NO: 172). In some embodiments, the variant STAT binding motif includes YLPL (SEQ ID NO: 173). In some embodiments, the variant STAT binding motif includes YLKQ (SEQ ID NO: 174).

[0053] In some embodiments, the variant IL-9R intracellular domain or its variant contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:107, SEQ ID NO:108, or SEQ ID NO:109. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence of SEQ ID NO:107, SEQ ID NO:108, or SEQ ID NO:109. In some embodiments, the chimeric JAK / STAT fusion domain contains a JAK-binding domain derived from a type I cytokine receptor and a STAT-binding domain derived from the IL-9R intracellular domain.

[0054] In some embodiments, the IL-9R STAT-binding domain includes amino acid residues 73-230 of SEQ ID NO:8. In some embodiments, the STAT-binding domain is 59-158 amino acids long and contains an IL-9R STAT-binding motif. In some embodiments, the IL-9R STAT-binding motif contains YLPQ (SEQ ID NO:171). In some embodiments, the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking a continuous amino acid sequence at the N-terminus of SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking a continuous amino acid sequence at the C-terminus of SEQ ID NO:8.

[0055] In some embodiments, the IL-9R STAT-binding domain is a truncated IL-9R STAT-binding domain lacking amino acids at positions 1-72 and / or 132-230 of SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain contains an amino acid sequence that is at least approximately 85% identical to that of SEQ ID NO:122 or SEQ ID NO:123. In some embodiments, the IL-9R STAT-binding domain contains the amino acid sequence of SEQ ID NO:122 or SEQ ID NO:123.

[0056] In some embodiments, the type I cytokine receptor is selected from the group consisting of interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R), interleukin-7 receptor (IL-7R), interleukin-13 receptor (IL-13R), interleukin-15 receptor (IL-15R), and interleukin-2 receptor (IL-21R). In some embodiments, the type I cytokine receptor is IL-7R. In some embodiments, the IL-7R JAK-binding domain is 65 amino acids long and contains a box-1 motif. In some embodiments, the IL-7R JAK-binding domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:121. In some embodiments, the IL-7R JAK-binding domain contains the amino acid sequence of SEQ ID NO:121.

[0057] In some embodiments, the chimeric JAK / STAT fusion domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:114 or SEQ ID NO:116.

[0058] In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:113, SEQ ID NO:115, or SEQ ID NO:117.

[0059] In some embodiments, synthetic cytokine receptors are constitutively active cytokine receptors. In some embodiments, synthetic cytokine receptors induce signaling through the STAT1, STAT3, and / or STAT5 pathways. In some embodiments, signaling through STAT1, STAT3, and / or STAT5 is increased compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R. In some embodiments, signaling through STAT1, STAT3, and / or STAT5 is sustained over a longer period compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R. In some embodiments, sustained STAT1, STAT3, and / or STAT5 signaling is determined by the phosphorylation state of STAT1, STAT3, and / or STAT5. In some aspects, polynucleotides encoding any of the synthetic cytokine receptors provided herein are provided herein.

[0060] In some aspects, vectors comprising any of the polynucleotides provided herein are provided herein.

[0061] In some aspects, methods for modifying isolated cells are provided herein, comprising the step of contacting the cells with any of the polynucleotides or vectors provided herein. In some aspects, modified cells expressing any of the synthetic cytokine receptors provided herein are provided herein.

[0062] In some aspects, synthetic cytokine receptors are provided herein that are homodimers of the same polypeptide chain, each comprising an extracellular domain, a transmembrane domain, and an intracellular domain capable of interleukin-9 receptor (IL-9R) signaling. In some embodiments, the IL-9R signaling intracellular domain comprises an IL-9R intracellular domain or a variant thereof. In some embodiments, the IL-9R signaling intracellular domain comprises a chimeric JAK / STAT fusion domain.

[0063] Modified cells expressing a synthetic cytokine receptor, which is a constitutively active cytokine receptor comprising an extracellular domain, a transmembrane domain, and an intracellular domain of the interleukin-9 receptor (IL-9R) or a variant thereof, are provided herein.

[0064] In some embodiments, the synthetic cytokine receptor is a multimer. In some embodiments, the synthetic cytokine receptor is a multimer of identical polypeptide chains, each containing an extracellular domain, a transmembrane domain, and an IL-9R intracellular domain or a variant thereof. In some embodiments, the multimer is a dimer. In some embodiments, the dimer is a homodimer. In some embodiments, each polypeptide chain is constitutively multimerized.

[0065] In some embodiments, the synthetic cytokine receptor comprises at least one self-assembling domain. In some embodiments, the at least one self-assembling domain is an extracellular domain and / or a transmembrane domain. In some embodiments, the synthetic cytokine receptor is multimerized through the transmembrane domain and / or the extracellular domain.

[0066] In some embodiments, synthetic cytokine receptors are multimerized via transmembrane and extracellular domains.

[0067] In some aspects, modified cells expressing a synthetic cytokine receptor which is a homodimer of the same polypeptide chain comprising an extracellular domain, a transmembrane domain, and an intracellular domain of the interleukin-9 receptor (IL-9R) or a variant thereof are provided herein. In some embodiments, the extracellular domain and / or transmembrane domain are heterogeneous with respect to IL-9R. In some embodiments, the transmembrane domain and extracellular domain are derived from the same protein. In some embodiments, the transmembrane domain and extracellular domain are derived from different proteins.

[0068] In some embodiments, the transmembrane domain is 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 amino acids long. In some embodiments, the transmembrane domain includes a transmembrane domain derived from glycophorin A (GpA), carnitine palmitoyltransferase 1 (CPT1), tumor necrosis factor receptor (TNFR), or Muc24. In some embodiments, the transmembrane domain includes a transmembrane domain derived from the thrombopoietin receptor.

[0069] In some embodiments, the transmembrane domain promotes α-helix dimerization. In some embodiments, the transmembrane domain contains motif GXXXG (SEQ ID NO:68). In some embodiments, the transmembrane domain is a motif Includes TIFF2026525229000003.tif4128.

[0070] In some embodiments, the transmembrane domain is a transmembrane domain derived from glycophorin A (GpA), or a variant thereof containing one or more mutations (e.g., 1, 2, 3, 4, 5, or 6 mutations) compared to the wild-type GpA transmembrane domain, wherein the variant GpA is sufficient to promote α-helix dimerization. In some embodiments, the transmembrane domain comprises a transmembrane domain derived from glycophorin A (GpA).

[0071] In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:22 or SEQ ID NO:43.

[0072] In some embodiments, the transmembrane domain includes a GXXXG (SEQ ID NO:68) motif and a GXXXA (SEQ ID NO:69) motif. In some embodiments, the transmembrane domain includes a transmembrane domain derived from carnitine palmitoyltransferase 1 (CPT1). In some embodiments, the transmembrane domain includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:45. In some embodiments, the transmembrane domain includes the amino acid sequence of SEQ ID NO:45.

[0073] In some embodiments, the synthetic cytokine receptor comprises motif AXXXA (SEQ ID NO: 77) or AXXXS (SEQ ID NO: 78). In some embodiments, the synthetic cytokine receptor comprises motif ΦPXΦ (SEQ ID NO: 75) or ΦTXXAΦ (SEQ ID NO: 76). In some embodiments, the transmembrane domain is derived from TNFR. In some embodiments, TNFR is TACI, DR5, p75NTR, Fas, TNFR1, TNFR2, or OX40.

[0074] In some embodiments, the transmembrane domain includes a transmembrane domain derived from DR5. In some embodiments, the transmembrane domain includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:46. In some embodiments, the transmembrane domain includes the amino acid sequence of SEQ ID NO:46.

[0075] In some embodiments, the transmembrane domain includes a transmembrane domain derived from TACI. In some embodiments, the transmembrane domain includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:44. In some embodiments, the transmembrane domain includes the amino acid sequence of SEQ ID NO:44.

[0076] In some embodiments, the transmembrane domain contains 1 to 6 cysteine ​​residues. In some embodiments, the transmembrane domain promotes disulfide dimerization. In some embodiments, disulfide dimerization forms 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

[0077] In some embodiments, the transmembrane domain is a variant transmembrane domain containing one or more mutations compared to the wild-type transmembrane domain to promote homodimerization of the receptor polypeptide. In some embodiments, one or more mutations promote α-helix dimerization or disulfide bond dimerization.

[0078] In some embodiments, one or more mutations introduce at least one cysteine ​​into the transmembrane domain. In some embodiments, one or more mutations introduce proline into the transmembrane domain. In some embodiments, one or more mutations introduce threonine into the transmembrane domain. In some embodiments, one or more mutations introduce a trimer peptide of cysteine, proline, and another amino acid other than cysteine ​​and proline into the transmembrane domain. In some embodiments, one or more mutations introduce a trimer peptide (CPT or TCP) of cysteine, proline, and threonine into the transmembrane domain.

[0079] In some embodiments, the transmembrane domain is a variant IL-7R transmembrane domain, and one or more mutations are in the wild-type transmembrane sequence. It is located within TIFF2026525229000004.tif4128.

[0080] In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:21.

[0081] In some embodiments, the transmembrane domain includes a transmembrane domain derived from Muc24. In some embodiments, the transmembrane domain includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:23. In some embodiments, the transmembrane domain includes the amino acid sequence of SEQ ID NO:23.

[0082] In some embodiments, the extracellular domain is approximately 150–260 amino acids long. In some embodiments, the extracellular domain is a dimerization domain. In some embodiments, the dimerization domain includes a hinge region. In some embodiments, the extracellular domain promotes disulfide bond dimerization. In some embodiments, the extracellular domain contains 1–6 cysteine ​​residues. In some embodiments, disulfide bond dimerization forms 1–4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

[0083] In some embodiments, the extracellular domain is derived from the extracellular domain of CD34, DAP12, glycophorin A, CD8, or Muc24. In some embodiments, the extracellular domain includes an extracellular domain derived from the thrombopoietin receptor.

[0084] In some embodiments, the extracellular domain comprises the extracellular domain of CD8, or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain comprises an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:18 or SEQ ID NO:19. In some embodiments, the extracellular domain comprises the amino acid sequence of SEQ ID NO:18 or SEQ ID NO:19.

[0085] In some embodiments, the extracellular domain comprises the extracellular domain of CD34, or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain comprises an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:4. In some embodiments, the extracellular domain comprises the amino acid sequence of SEQ ID NO:4.

[0086] In some embodiments, the extracellular domain is the extracellular domain of Muc24, or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:20. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:20. In some embodiments, the extracellular domain is the extracellular domain of DAP12, or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:5. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:5.

[0087] In some embodiments, the extracellular domain is the extracellular domain of glycophorin A (GpA), or a truncated portion thereof containing at least one cysteine ​​residue. In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:16 or SEQ ID NO:17. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:16 or SEQ ID NO:17.

[0088] In some embodiments, the IL-9R intracellular domain or its variant is approximately 100 to 260 amino acids long. In some embodiments, the IL-9R intracellular domain or its variant contains a box 1 motif and / or a box 2 motif. In some embodiments, the IL-9R intracellular domain or its variant contains a box 2 motif. In some embodiments, the IL-9R intracellular domain or its variant is 230 amino acids long.

[0089] In some embodiments, the IL-9R intracellular domain is either the wild-type IL-9R intracellular domain or a variant thereof containing one or more mutations compared to the wild-type IL-9R intracellular domain shown in SEQ ID NO:8.

[0090] In some embodiments, one or more mutations include one or more amino acid insertions, deletions, and / or substitutions. In some embodiments, one or more mutations facilitate signaling through the STAT1 pathway, the STAT3 pathway, and / or the STAT5 pathway.

[0091] In some embodiments, the IL-9R intracellular domain or its variants contain an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain or its variants contain the amino acid sequence of SEQ ID NO:8.

[0092] In some embodiments, the IL-9R intracellular domain or its variant contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, or SEQ ID NO:56.

[0093] In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:67. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence of SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:67.

[0094] In some embodiments, the IL-9R intracellular domain or its variants contain one or more amino acid deletions relative to the wild-type IL-9R intracellular domain (SEQ ID NO: 8). In some embodiments, the IL-9R intracellular domain or its variants are truncated IL-9R lacking the continuous amino acid sequence at the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the truncated IL-9R intracellular domain or its variants are shortened by only 62 to 99 consecutive amino acids from the C-terminus of the wild-type IL-9R intracellular domain.

[0095] In some embodiments, the intracellular domain of IL-9R or its variants are a shortened form of IL-9R, lacking amino acids 132-230 of SEQ ID NO:8 or amino acids 134-230 of SEQ ID NO:8.

[0096] In some embodiments, the IL-9R intracellular domain or its variants contain an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:54, SEQ ID NO:103, or SEQ ID NO:104.

[0097] In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, or SEQ ID NO:63.

[0098] In some embodiments, the IL-9R intracellular domain or its variants contain one or more amino acid substitutions relative to the wild-type IL-9R intracellular domain (SEQ ID NO:8).

[0099] In some embodiments, the IL-9R intracellular domain or a variant thereof includes a STAT-binding motif or a variant thereof. In some embodiments, the STAT-binding motif includes a STAT1-binding motif, a STAT3-binding motif, and / or a STAT5-binding motif. In some embodiments, the STAT-binding motif includes YLPQ (SEQ ID NO: 171). In some embodiments, the STAT-binding motif includes a variant STAT-binding motif. In some embodiments, the variant STAT-binding motif includes YRPQ (SEQ ID NO: 172). In some embodiments, the variant STAT-binding motif includes YLPL (SEQ ID NO: 173). In some embodiments, the variant STAT-binding motif includes YLKQ (SEQ ID NO: 174). In some embodiments, the variant IL-9R intracellular domain or a variant thereof includes an amino acid sequence that is at least approximately 85% identical to SEQ ID NO: 107, SEQ ID NO: 108, or SEQ ID NO: 109.

[0100] In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence of SEQ ID NO:107, SEQ ID NO:108, or SEQ ID NO:109.

[0101] In some embodiments, the chimeric JAK / STAT fusion domain comprises a JAK-binding domain derived from a type I cytokine receptor and a STAT-binding domain derived from an IL-9R intracellular domain.

[0102] In some embodiments, the IL-9R STAT-binding domain includes amino acid residues 73-230 of SEQ ID NO:8.

[0103] In some embodiments, the STAT-binding domain is 59-158 amino acids long and contains an IL-9R STAT-binding motif. In some embodiments, the IL-9R STAT-binding motif contains YLPQ (SEQ ID NO: 171). In some embodiments, the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking a continuous amino acid sequence at the N-terminus of SEQ ID NO: 8. In some embodiments, the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking a continuous amino acid sequence at the C-terminus of SEQ ID NO: 8.

[0104] In some embodiments, the IL-9R STAT-binding domain is a truncated IL-9R STAT-binding domain lacking amino acids at positions 1-72 and / or 132-230 of SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain contains an amino acid sequence that is at least approximately 85% identical to that of SEQ ID NO:122 or SEQ ID NO:123. In some embodiments, the IL-9R STAT-binding domain contains the amino acid sequence of SEQ ID NO:122 or SEQ ID NO:123.

[0105] In some embodiments, the type I cytokine receptor is selected from the group consisting of interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R), interleukin-7 receptor (IL-7R), interleukin-13 receptor (IL-13R), interleukin-15 receptor (IL-15R), and interleukin-2 receptor (IL-21R). In some embodiments, the type I cytokine receptor is IL-7R.

[0106] In some embodiments, the IL-7R JAK-binding domain is 65 amino acids long and contains a box-1 motif. In some embodiments, the IL-7R JAK-binding domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:121. In some embodiments, the IL-7R JAK-binding domain contains the amino acid sequence of SEQ ID NO:121. In some embodiments, the chimeric JAK / STAT fusion domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:114 or SEQ ID NO:116. In some embodiments, the chimeric JAK / STAT fusion domain contains the amino acid sequence of SEQ ID NO:114 or SEQ ID NO:116.

[0107] In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:113, SEQ ID NO:115, or SEQ ID NO:117.

[0108] In some embodiments, synthetic cytokine receptors are constitutively active cytokine receptors.

[0109] In some embodiments, synthetic cytokine receptors induce signaling through the STAT1, STAT3, and / or STAT5 pathways. In some embodiments, signaling through STAT1, STAT3, and / or STAT5 is increased compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R. In some embodiments, signaling through STAT1, STAT3, and / or STAT5 is sustained over longer periods compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R. In some embodiments, sustained STAT1, STAT3, and / or STAT5 signaling is determined by the phosphorylation state of STAT1, STAT3, and / or STAT5.

[0110] In some embodiments, the modified cells further express at least one distinct type of modified receptor. In some embodiments, the at least one distinct type of modified receptor is a chimeric antigen receptor. In some embodiments, the extracellular domain of the chimeric antigen receptor binds to an antigen expressed in cancer cells. In some embodiments, the extracellular domain of the chimeric antigen receptor binds to an idiotype of an antibody. In some embodiments, the antibody is against an antigen expressed in cancer cells. In some embodiments, the cancer cells are hematological cancer cells or solid tumor cancer cells.

[0111] In some aspects, the cells are immune cells. In some aspects, the cells are lymphocytes. In some aspects, the modified cells are immune effector cells. In some aspects, the cells are T cells or natural killer (NK) cells. In some aspects, the cells are T cells, and the T cells are CD4+ T cells or CD8+ T cells. In some aspects, the immune effector cells are cytotoxic T cells. In some aspects, the immune effector cells are natural killer cells. In some aspects, the cells are primary cells. In some aspects, the cells are human cells. In some aspects, a cell population comprising at least one of the modified cells provided herein is provided herein. In some aspects, at least one modified cell comprises modified CD4+ T cells and modified CD8+ T cells.

[0112] In some aspects, pharmaceutical compositions comprising any of the modified cells or any of the cell populations provided herein are provided herein. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition further comprises a cryoprotective substance. In some embodiments, the pharmaceutical composition is intended for use in the treatment of cancer in a subject.

[0113] In some aspects, methods for treating a disease or condition in a subject are provided herein, comprising the step of administering a therapeutically effective amount of any pharmaceutical composition provided herein to the subject. In some embodiments, the disease or condition is cancer.

[0114] In some embodiments, modified cells of the pharmaceutical composition express modified antigen receptors that bind to antigens expressed in cancer cells. In some embodiments, the modified antigen receptor is a chimeric antigen receptor. In some embodiments, the modified antigen receptor is a T cell receptor.

[0115] In some aspects, methods for improving the function of immune cells are provided herein, comprising the step of introducing one of the polynucleotides or vectors provided herein into immune cells, wherein the improvement of immune cell function includes an increase in STAT1 signaling, STAT3 signaling, and / or STAT5 signaling compared to immune cells expressing wild-type IL-9R.

[0116] In some aspects, methods for improving the cytotoxicity of immune cells are provided herein, comprising the step of introducing one of the polynucleotides or vectors provided herein into immune cells, wherein the improvement in the cytotoxicity of immune cells includes increased target cell killing compared to immune cells expressing wild-type IL-9R.

[0117] In some aspects, methods for improving the viability of immune cells are provided herein, comprising the step of introducing one of the polynucleotides or vectors provided herein into immune cells, wherein the improvement in immune cell viability includes a reduction in immune cell death compared to immune cells expressing wild-type IL-9R.

[0118] In some aspects, methods for improving cytokine secretion by immune cells are provided herein, comprising the step of introducing one of the polynucleotides or vectors provided herein into immune cells, wherein the improvement in cytokine secretion includes an increase in interferon secretion compared to immune cells expressing wild-type IL-9R.

[0119] In some embodiments, the interferon comprises IFN-γ. In some embodiments, the immune cells further comprise a modified antigen receptor. In some embodiments, one distinct type of modified receptor is a chimeric antigen receptor (CAR). In some embodiments, the extracellular domain of the CAR binds to an antigen expressed in cancer cells. In some embodiments, the cancer cells are hematological cancer cells or solid tumor cancer cells.

[0120] In some aspects, immune cells are lymphocytes. In some aspects, immune cells are effector cells. In some aspects, immune cells are T cells or NK cells. In some aspects, immune cells are T cells. In some aspects, immune cells are T cells, and the T cells are CD4+ T cells or CD8+ T cells. In some aspects, immune cells are cytotoxic T cells. In some aspects, immune cells are NK cells. In some aspects, immune cells are primary cells.

[0121] In some embodiments, methods for improving the function of immune cells are provided herein, which include the step of introducing one of the polynucleotides or vectors provided herein into immune cells, thereby improving the function of the immune cells.

[0122] In some embodiments, improvements in immune cell function include one or more of the following compared to reference cells: increased STAT signaling, increased cytotoxicity, increased proliferation, increased viability, increased cytokine secretion, and increased cytotoxic protein secretion.

[0123] In some embodiments, reference cells include unmodified or modified immune cells. In some embodiments, modified immune cells express modified antigen receptors. In some embodiments, the modified antigen receptor is wild-type IL-9R, a chimeric antigen receptor (CAR), or a T cell receptor (TCR).

[0124] In some embodiments, increased STAT signaling includes increased STAT1 signaling, STAT3 signaling, and / or STAT5 signaling. In some embodiments, increased cytotoxicity includes increased killing of target cells. In some embodiments, increased cytokine secretion includes increased secretion of one or more of interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-10 (IL-10), and TNF-α. In some embodiments, increased cytotoxic protein secretion includes increased secretion of one or more of granzyme A, granzyme B, granulysin, and perforin. In some embodiments, increased viability includes decreased cell death.

[0125] In some aspects, immune cells are lymphocytes. In some aspects, immune cells are effector cells. In some aspects, immune cells are T cells or NK cells. In some aspects, immune cells are T cells, and these T cells are CD4+ T cells or CD8+ T cells. In some aspects, immune cells are cytotoxic T cells. In some aspects, immune cells are NK cells. In some aspects, immune cells are primary cells. [Brief explanation of the drawing]

[0126] [Figure 1] This shows an exemplary design of a synthetic cytokine receptor. [Figure 2] This image shows exemplary synthetic cytokine receptor expression on the surface of primary human CD3-positive T cells, including CD8+ T cells (left panel) or CD4+ T cells (right panel). [Figure 3A]Figures 3A and 3B show the surface expression levels of CD27 and CD45RA in primary human CD3-positive T cells transduced 5 days after transduction with a lentiviral vector expressing the 9RC synthetic cytokine receptor, or in untransduced primary human CD3-positive T cells ("UTD"). Figure 3A is an exemplary flow cytometry dot plot with CD45RA staining on the x-axis and CD27 staining on the y-axis. Figure 3B illustrates the percentage of CD45RA-positive and CD27-positive T cells in UTD T cells or 9RC+ transduced T cells. [Figure 3B] Please refer to the explanation in Figure 3A. [Figure 4] This shows the surface expression levels of CD27 and CD45RA in primary human CD3-positive T cells 12 days after transduction with a lentiviral vector expressing exemplary synthetic cytokine receptors (9RC, 9RC2, or 9RD2), or in primary human CD3-positive T cells that were not transduced. [Figure 5] This shows the surface expression levels of CD27 and Fas 16 days after transduction of primary human CD3-positive T cells using a lentiviral vector expressing exemplary synthetic cytokine receptors (9RC, 9RC2, or 9RD2), or in primary human CD3-positive T cells that were not transduced. [Figure 6A] This shows the surface expression levels of CD27 and Fas in all cells, or untransduced cells, 16 days after transduction of primary human CD3-positive T cells with a lentiviral vector expressing an exemplary synthetic cytokine receptor (9RC or 9RC2), specifically the exemplary synthetic cytokine receptor 9RC co-expressed with a CAR isolated by a 2A sequence (named "40bbz-2a-CD34-9RC"). [Figure 6B]The surface expression levels of CD27 and Fas in Flag-positive cells 16 days after transduction of primary human CD3-positive T cells with a lentiviral vector expressing an exemplary synthetic cytokine receptor (9RC or 9RC2), or an exemplary synthetic cytokine receptor 9RC co-expressed with a CAR isolated by a 2A cleavable linker (named "40bbz-2a-CD34-9RC"). [Figure 7A] This shows the surface expression levels of CCR7 and CD45RA in all cells or untransduced cells 16 days after transduction of primary human CD3-positive T cells with a lentiviral vector expressing an exemplary synthetic cytokine receptor (9RC or 9RC2), co-expressed with a CAR isolated by a 2A cleavable linker (named "40bbz-2a-CD34-9RC"). [Figure 7B] The surface expression levels of CCR7 and CD45RA in Flag-positive cells 16 days after transduction of primary human CD3-positive T cells with a lentiviral vector expressing an exemplary synthetic cytokine receptor (9RC or 9RC2), or an exemplary synthetic cytokine receptor 9RC co-expressed with a CAR isolated by a 2A cleavable linker (named "40bbz-2a-CD34-9RC"). [Figure 8] This shows the surface expression level of CD27 in 9RC-expressing primary human CD3-positive T cells based on geometric mean fluorescence intensity (gMFI) measured by flow cytometry. [Figure 9A]Figures 9A-9B show the surface expression levels of CD57 in 9RC-expressing primary human CD3-positive T cells, determined by co-staining with a FLAG tag after transduction with a lentiviral vector expressing the 9RC synthetic cytokine receptor, or in primary human CD3-positive T cells that were not transduced ("UTD"). Figure 9A is a representative flow cytometry dot plot with CD57 staining on the y-axis and FLAG staining on the x-axis. Figure 9B illustrates the percentage of CD57-high-expressing (CD57hi) CD8+ T cells in UTD T cells or 9RC+ transduced T cells. [Figure 9B] Please refer to the explanation in Figure 9A. [Figure 10] This shows the surface expression levels of TIM3 and LAG3 in primary human CD3-positive T cells co-transduced with lentiviral vectors expressing 9RC synthetic cytokine receptor and CAR isolated by either a 2A sequence (40bbz-2a-FLAG-9RC) or an IRES sequence (40bbz-IRES-CD34-9RC), or in primary human CD3-positive T cells that were not transduced ("UTD"). An exemplary flow cytometry dot plot is shown with TIM3 staining on the y-axis and LAG3 staining on the x-axis. [Figure 11] This shows the surface expression levels of CD4 and CD8 in primary human CD3-positive T cells 5 days after transduction with a lentiviral vector expressing exemplary synthetic cytokine receptors (9RC, 9RC2, or 9RD2), or in primary human CD3-positive T cells that were not transduced. [Figure 12] This shows the proliferation of primary human CD3-positive T cells expressing exemplary synthetic cytokine receptors (9RC or 9RC2), exemplary synthetic cytokine receptor 9RC co-expressed with a CAR (named "40bbz-9RC"), or untransduced (UTD) cells in long-term in vitro culture. The results illustrate the culture days after activation and the multiplicative increase in cell count compared to the previous count for each condition. [Figure 13A]Figures 13A–13E show the design and expression of additional exemplary synthetic cytokine receptors, including IL7R TMD and IL-9R ICD. Each of the additional synthetic receptor constructs had one of the following distinct extracellular domains (ECD): GpA (construct “G-7-9R”; Figure 13A), truncated GpA (construct “tG-7-9R”; Figure 13B), CD8 (construct “CD8-7-9R”; Figure 13C), truncated CD8 (construct “tCD8-7-9R”; Figure 13D), or Muc24 (construct “M-7-9R”; Figure 13E). [Figure 13B] Please refer to the explanation in Figure 13A. [Figure 13C] Please refer to the explanation in Figure 13A. [Figure 13D] Please refer to the explanation in Figure 13A. [Figure 13E] Please refer to the explanation in Figure 13A. [Figure 14A] Figures 14A and 14B show the design and expression of additional exemplary synthetic cytokine receptors containing transmembrane domains other than the IL-9R ICD and IL7R TMD. Figure 14A illustrates a synthetic cytokine construct ("GG-9R") containing glycophorin A (GpA) ECD, GpA TMD, and IL-9R ICD. Figure 14B illustrates a synthetic cytokine receptor ("MM-9R") having Muc24 ECD, Muc24 TMD, and IL-9R ICD. [Figure 14B] Please refer to the explanation in Figure 14A. [Figure 15A]Figures 15A–15H show design and expression data for additional exemplary synthetic cytokine receptors, including IL9R ICD, full-length or truncated ECD derived from GpA, CD8, or Muc24, and TMD derived from GpA, truncated GpA (named GpA*), TACI, CPT1, or DR5. The illustrated structures include the following ECDs, TMDs, and ICDs: abbreviated GpA ECD, GpA TMD, and IL9R ICD (structure "tGpA-G-9R"; Figure 15A), CD8 ECD, GpA TMD, and IL9R ICD (structure "CD8-G-9R"; Figure 15B), abbreviated CD8 ECD, GpA TMD, and IL9R ICD (structure "tCD8-G-9R"; Figure 15C), Muc24 ECD, GpA TMD, and IL9R ICD (structure "M24-G-9R"; Figure 15D), abbreviated CD8 ECD, abbreviated GpA TMD, and IL9R ICD (structure "tCD8-G*-9R"; Figure 15E), abbreviated CD7 ECD, TACI TMD, and IL9R ICD (construction "tCD8-T-9R"; Figure 15F), abbreviated CD8 ECD, CPT1 TMD, and IL9R ICD (construction "tCD8-C-9R"; Figure 15G), as well as abbreviated CD8 ECD, DR5 TMD, and IL9R ICD (construction "tCD8-D-9R"; Figure 15H). [Figure 15B] Please refer to the explanation in Figure 15A. [Figure 15C] Please refer to the explanation in Figure 15A. [Figure 15D] Please refer to the explanation in Figure 15A. [Figure 15E] Please refer to the explanation in Figure 15A. [Figure 15F] Please refer to the explanation in Figure 15A. [Figure 15G] Please refer to the explanation in Figure 15A. [Figure 15H] Please refer to the explanation in Figure 15A. [Figure 16]This study shows the transduction efficiency of exemplary synthetic cytokine receptors in primary human CD3-positive T cells (UTDs) 5 days after transduction with a lentiviral vector expressing exemplary synthetic cytokine receptors, or in T cells that were not transduced. Transduction efficiency was determined by flow cytometry using cell surface staining for Flag-tag-positive cells. [Figure 17A] Figures 17A–17C show the surface expression levels of CD27 and CD45RA five days after transduction of primary human CD3-positive T cells with a lentiviral vector expressing exemplary synthetic cytokine receptors. The figures provide representative flow cytometry contour plots with CD45RA staining on the x-axis and CD27 staining on the y-axis. [Figure 17B] Please refer to the explanation in Figure 17A. [Figure 17C] Please refer to the explanation in Figure 17A. [Figure 18] This shows the in vivo survival rate of tumor-carrying mice treated with primary human CD3-positive T cells co-expressing CAR(40BBz) and exemplary synthetic cytokine receptors, compared to primary human CD3-positive T cells expressing CAR alone or untransduced (UTD) primary human CD3-positive T cells. [Figure 19] This shows STAT phosphorylation (pSTAT) in primary human CD3-positive T cells expressing exemplary synthetic cytokine receptors. pSTAT was measured at 0 hours, 20 minutes, 2 hours, and 24 hours after cytokine stimulation. [Figure 20] This shows exemplary expression of synthetic cytokine receptors on the surface of primary human CD3-positive T cells. [Figure 21] This shows STAT phosphorylation (pSTAT) of STAT1, STAT3, and STAT5 in primary human CD3-positive T cells expressing exemplary synthetic cytokine receptors. [Figure 22] This shows the number of HCT116 tumor cells per field of view (FOV) over time in a T cell-mediated killing assay. T cells were obtained from two healthy donors and co-cultured with tumor cells in an effector:target (E:T) ratio of 1:2. [Figure 23] This shows the number of HCT116 tumor cells per field of view (FOV) over time in a T cell-mediated killing assay. T cells were obtained from one healthy donor and co-cultured with tumor cells in an effector:target (E:T) ratio of 1:4. [Figure 24] This shows the phenotypic markers CD27 and CD45RA in primary T cells expressing exemplary synthetic cytokine receptors. [Figure 25] This shows the total number of T cells co-expressing CAR(40BBz) and synthetic cytokine receptors at the time of seeding and after two rounds of T cell-mediated killing. [Figure 26] This shows the percentage of cytokine-positive CD8 T cells after targeted stimulation. CD8 T cells co-expressed CAR(40BBz) and exemplary synthetic cytokine receptors, or expressed CAR alone. Measured cytokines included IL-2, IFN-γ, and TNF-α. [Figure 27] This shows the percentage of surviving T cells after two rounds of stimulation. T cells co-expressing CAR(40BBz) and a synthetic cytokine receptor were co-cultured with K562 cells expressing an antigen recognized by CAR(40BBz). [Figure 28A] Figures 28A-28B show the transduction percentages of T cells expressing CAR(40BBz) and exemplary synthetic cytokine receptors. 9RC5=C-7-9R(a); 9RC5.1=CD8-7-9R(c); 9RC5.2=tCD8-7-9R(c). [Figure 28B] Please refer to the explanation in Figure 28A. [Figure 29] This image shows STAT phosphorylation (pSTAT) in primary human CD3-positive T cells co-expressing CAR(40BBz) and an exemplary synthetic cytokine receptor. pSTAT1, pSTAT3, and pSTAT5 are expressed as percentages of the highest signal observed for each individual STAT. [Figure 30]This shows the percentage of CD8+ T cells expressing cytokines after 4 hours of stimulation by target cells. CD8+ T cells either co-expressed CAR(40BBz) and exemplary synthetic cytokine receptors, or expressed CAR alone. Measured cytokines included IFN-γ, IL-2, IL-10, and TNF-α. [Figure 31] This shows the secretion of cytokines and effector molecules by CD8+ T cells 24 hours after stimulation by target cells. CD8+ T cells either co-expressed CAR(40BBz) and an exemplary synthetic cytokine receptor, or expressed CAR alone. Secretion of cytokines and effector molecules is expressed as a percentage of the peak expression of each analyte. [Figure 32A] Figures 32A–32B show the staining of CD45RA and CD27 in CD8+ T cells in serial co-culture with target cells. CD8+ T cells co-expressed CAR(40BBz) and exemplary synthetic cytokine receptors, or expressed CAR alone. Figure 32A illustrates the expression of CD45RA and CD27 over 3 weeks. Figure 32B illustrates summary data from three technical iterations at 3 weeks. [Figure 32B] Please refer to the explanation in Figure 32A. [Figure 33A] Figures 33A-33C show the number of viable CD8+ T cells in continuous co-culture with target cells. CD8+ T cells either co-expressed CAR(40BBz) and an exemplary synthetic cytokine receptor, or expressed CAR only. Figure 33A illustrates the number of viable CD8+ T cells over 3 weeks. Figure 33B illustrates the number of viable CD8+ T cells after 3 weeks. Figure 33C illustrates CD39 expression in CD8+ T cells after 3 weeks. [Figure 33B] Please refer to the explanation in Figure 33A. [Figure 33C] Please refer to the explanation in Figure 33A. [Figure 34]The images show an increase in CD8+ T cells cultured for 4 weeks with and without IL-2. CD8+ T cells either co-expressed CAR(40BBz) and exemplary synthetic cytokine receptors, or expressed CAR alone. [Figure 35] This shows tumor growth in tumor-carrying mice administered with primary human CD3-positive T cells co-expressing CAR(40BBz) and an exemplary synthetic cytokine receptor, compared to primary human CD3-positive T cells expressing CAR alone or untransduced (UTD). Tumor-carrying mice were administered 1 × 10⁶ cells. [Figure 36] This shows tumor growth in tumor-carrying mice administered with primary human CD3-positive T cells co-expressing CAR(40BBz) and exemplary synthetic cytokine receptors, compared to primary human CD3-positive T cells expressing CAR alone or untransduced (UTD). Tumor-carrying mice were administered 3 × 10⁵ cells. [Modes for carrying out the invention]

[0127] Detailed explanation This disclosure provides synthetic cytokine receptors that can drive interleukin-9 (IL-9) receptor (IL-9R) signaling even in the absence of congeneral cytokine ligands (i.e., constitutively active). These synthetic cytokine receptors include an intracellular domain derived from the intracellular domain of human IL-9 receptor α (IL-9Rα) that can activate STAT1, STAT3, and STAT5 and induce a potent JAK / STAT signaling cascade. These synthetic cytokine receptors also include various modified intracellular, transmembrane, and extracellular domains. Immune cells expressing these synthetic cytokine receptors, such as T cells, are advantageous because such T cells exhibit the characteristics of stem memory T cells without showing signs of malignant transformation. Such T cells may be used in adoptive immunotherapy.

[0128] In some embodiments, the mechanism of constitutive activation includes multimerization of the extracellular domain. In some embodiments, the mechanism of constitutive activation includes multimerization of the transmembrane domain. In some embodiments, the mechanism of constitutive activation includes multimerization of the intracellular domain. In some embodiments, the mechanism of constitutive activation includes multimerization of at least two domains (e.g., an extracellular domain and a transmembrane domain). In some embodiments, the mechanism of constitutive activation includes multimerization of three domains (i.e., an extracellular domain, a transmembrane domain, and an intracellular domain).

[0129] Adoptive-transplanted genetically modified immune cells (e.g., T cells) exhibit considerable antitumor activity in patients with hematopoietic malignancies, but have limited efficacy in solid tumors. This is due to the immunosuppressive environment of solid tumors, inefficient tumor invasion, and chronic antigenic stimulation, which result in a lack of persistence and / or efficacy. In some cases, pluripotency and replication capacity are also reduced. Therefore, compositions and / or methods are needed to mitigate or eliminate immune cell exhaustion, lack of persistence, and / or reduced efficacy.

[0130] The disclosures provided address these needs. This disclosure demonstrates that the provided synthetic chimeric IL-9 receptors have the ability to constitutively activate cells expressing them, such as T cells, even in the absence of ligands. In some embodiments, this activation can induce a potent JAK / STAT signaling cascade, with STAT1, STAT3, and STAT5 all being activated. In some embodiments, this activation has the ability to lead to T cells that overcome T cell exhaustion by shifting T cells to a more naive phenotype and tilting the CD4:CD8 ratio to, for example, 2:1, without inducing malignant transformation of T cells. Thus, this disclosure demonstrates that the constitutive IL-9Rs disclosed herein have the ability to reprogram immune cells and alter their phenotype to overcome exhaustion, thereby improving antitumor activity in solid tumors.

[0131] I. Definition Unless otherwise defined herein, technical and scientific terms used herein have the meanings generally understood by those skilled in the art. For the interpretation of this specification, the following definitions of terms apply, and wherever the context indicates otherwise, the singular form of a term is also included in its plural form, and vice versa. In the event of any conflict between the definitions of terms provided herein and any documents incorporated herein by reference, the following definitions shall prevail.

[0132] The terms “a,” “an,” and “the” are plural in nature, as used herein, unless the context explicitly indicates otherwise.

[0133] The term "approximately" is understood to mean, as used herein, the number stated and ±10% of that number, or, in the case of a range, the number 10% below the stated lower limit and 10% above the stated upper limit for the stated value.

[0134] The terms "or" and "and / or" include any combination of one or more of the items described herein, as used herein.

[0135] The terms "including," "includes," "included," and other forms are not limited to those used herein.

[0136] "Comprise" and its grammatical equivalents, as used herein, specify the presence of the described features, integers, processes, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, processes, operations, elements, components, and / or groups thereof.

[0137] "Administer," "administer," or "to administer" means, as used herein, the act of injecting or otherwise physically delivering a substance (e.g., a pharmaceutical composition provided herein) into a subject (e.g., a human) by, for example, oral, mucosal, topical, intradermal, parenteral, intravenous, intravitreous, intra-articular, subretinal, intramuscular, subarachnoid space, and / or any other physical delivery method described herein or known in the art. Delivery may be systemic or to specific tissues.

[0138] The terms “to bind” or “to bond” refer to covalent or non-covalent interactions between molecules (e.g., the formation of complexes by interactions) as used herein. Exemplary non-covalent interactions include hydrogen bonds, ionic bonds, hydrophobic interactions, and / or van der Waals interactions. With regard to terms such as “specific binding,” “specifically binding,” or “specific to,” a binding is measurably different from a non-specific interaction. Specific binding can be measured, for example, by determining the binding of a molecule compared to the binding of a control molecule, the control molecule being typically a molecule with a similar structure but lacking binding activity.

[0139] The terms “chimeric antigen receptor” or “CAR” refer to a genetically modified receptor that can be used to transfer one or more antigen specificities to immune effector cells, such as T cells and NK cells, as used herein. CARs are also known as “artificial T cell receptors,” “chimeric T cell receptors,” or “chimeric immune receptors.” “CAR-T cells” refer to T cells that express a CAR. “CAR-NK cells” refer to NK cells that express a CAR.

[0140] The term "constitutively active" as used in relation to receptors refers to receptors that can activate downstream signaling (e.g., interleukin-9 signaling) even in the absence of their homologous ligands.

[0141] The term “contact” as used in relation to contact between target cells and a compound or other cells shall include incubation of the target cells and the compound or other cells together.

[0142] The terms “effective dose” or “therapeutic effective dose” mean, as used herein, an amount of therapeutic agent (e.g., a pharmaceutical composition provided herein) sufficient to treat, diagnose, prevent, delay the onset of, reduce the severity and / or duration of, and / or alleviate, a maximum of a given condition, disorder, or disease and / or symptoms associated therewith. This term also includes amounts necessary to reduce, slow, or alleviate the progression or progression of a given disease, to reduce, slow, or alleviate the recurrence, onset, or onset of a given disease, and / or to improve or enhance the preventive or therapeutic effect of another treatment, or to function as a bridge to another treatment.

[0143] The term “immune effector cells” as used herein refers to cells involved in initiating innate and adaptive immune responses, including but not limited to lymphocytes, natural killer (NK) cells, NKT cells, macrophages, monocytes, eosinophils, basophils, neutrophils, dendritic cells, and mast cells. In some embodiments, immune effector cells are lymphocytes. In some embodiments, immune effector cells are cytotoxic T cells, e.g., CD4+ T cells, CD8+ T cells (also called cytotoxic T cells or CTLs), regulatory T cells (Tregs), Th1 cells, Th2 cells, or Th17 cells.

[0144] The term "leucine zipper domain," as used herein, refers to an amphiphilic α-helix containing repeating Leu residues every seven residues on one face of the helix, and functions as a dimerization module. In some embodiments, the leucine zipper domain is an active leucine zipper domain. Upon dimerization with another leucine zipper domain, the leucine zipper α-helix forms a parallel coiled coil based on the packing of side chains at the hydrophobic interface. In some embodiments, the leucine zipper domain is an inactive leucine zipper domain that cannot dimerize with another leucine zipper domain.

[0145] The term “naturally occurring” as applied to nucleic acids, polypeptides, cells, or organisms refers to nucleic acids, polypeptides, cells, or organisms that are found in nature. For example, a polypeptide or polynucleotide sequence found in an organism (including viruses) that can be isolated from a natural origin and has not been intentionally modified by humans in the laboratory is considered naturally occurring.

[0146] The term “pharmaceutically acceptable excipient, carrier, or diluent” means, as used herein, any substance formulated together with the active ingredient of a pharmaceutical composition that enables the active ingredient to retain its biological activity and does not react with the target immune system. Such substances may be included for long-term stabilization, to excipient small amounts of solid formulations containing potent active ingredients, or to impart therapeutic enhancement to the active ingredient in the final dosage form, for example, to facilitate absorption, reduce viscosity, or increase solubility. The selection of an appropriate substance may depend on the route of administration and dosage form, as well as the active ingredient and other factors. Compositions containing such substances may be formulated by well-known conventional methods (see, for example, Remington, The Science and Practice of Pharmacy, 23rd edition, A. Adejare, ed, Academic Press, 2020).

[0147] The terms “pharmaceutical composition” or “therapeutic composition” mean, as used herein, a composition that can be administered to a subject for the treatment of a particular disease or disorder.

[0148] The terms “polynucleotide” or “nucleic acid” refer to deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and DNA / RNA hybrids as used herein. Polynucleotides may be single-stranded or double-stranded and may be recombinant, synthetic, or isolated. Polynucleotides include, but are not limited to, pre-messenger RNA (pre-mRNA), messenger RNA (mRNA), RNA, genomic DNA (gDNA), PCR-amplified DNA, complementary DNA (cDNA), synthetic DNA, or recombinant DNA. Polynucleotides may contain modified nucleotides or bases and / or analogues thereof, or any substrates that can be incorporated into a polymer by DNA polymerase or RNA polymerase, or by synthetic reactions. Unless otherwise specified, the left end of a single-stranded polynucleotide sequence disclosed herein is the 5' end; the left direction of a double-stranded polynucleotide sequence is referred to as the 5' direction. The direction of addition from 5' to 3' of a nascent RNA transcript is referred to as the transcription direction.

[0149] The terms “polypeptide,” “peptide,” and “protein” refer to polymers of amino acids of any length, as used herein. Polymers may be linear or branched, may contain modified amino acids, or may have non-amino acid intervening molecules. These terms also include naturally occurring or interveningly modified amino acid polymers (e.g., disulfide bond formation, glycosylation, lipidization, acetylation, phosphorylation, or other operations or modifications). For example, polypeptides containing one or more amino acid analogs, e.g., non-natural amino acids, and other modifications known in the art are also included in the definition.

[0150] As used herein, the term "population" of cells refers to any number of cells greater than 1, preferably at least about 1×10 3 cells, at least about 1×10 4 cells, at least about 1×10 5 cells, at least about 1×10 6 cells, at least about 1×10 7 cells, at least about 1×10 8 cells, at least about 1×10 9 cells, at least about 1×10 10 cells, at least about 1×10 11 cells, or more. A cell population can refer to an in vitro population (e.g., a population of cells in culture) or an in vivo population (e.g., a population of cells present in a particular tissue).

[0151] The term "sequence identity," as used herein, refers to the percentage of identical bases or amino acids in the same relative positions between the sequences of two polynucleotides or polypeptides. Thus, the sequence of one polynucleotide or polypeptide has a certain percentage of sequence identity compared to the sequence of the other polynucleotide or polypeptide. For sequence comparison, typically one sequence serves as a reference sequence, and the test sequence is compared to it. The term "reference sequence" refers to the molecule against which the test sequence is compared. Methods of sequence alignment for comparing and determining percent sequence identity and percent complementarity are well known in the art. Optimal alignment of sequences for comparison can be achieved using, for example, Needleman and Wunsch (1970) J.Mol.Biol.48:443 homology alignment algorithm, Pearson and Lipman (1988) Proc.Nat'l.Acad.Sci.USA 85:2444 similarity search method, computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA from the Wisconsin Genetics Software Package (Genetics Computer Group, 575 Science Dr., Madison, WI)), manual alignment and visual inspection (see, for example, Brent et al., (2003) Current Protocols in Molecular Biology), Altschul et al., (1977) Nuc.Acids Res.25:3389-3402; and Altschul et al. This can be carried out using algorithms known in the art, including the BLAST algorithm and the BLAST 2.0 algorithm described in al., (1990) J.Mol.Biol.215:403-410. Software for performing BLAST analysis is publicly available from the National Center for Biotechnology Information.To obtain nucleotide sequences homologous to the nucleic acid molecules described herein, a BLAST nucleotide search can be performed using, for example, XBLAST nucleotide program parameters set to a score of 100 and a word length of -2. To obtain amino acid sequences homologous to the protein molecules described herein, a BLAST protein search can be performed using, for example, XBLAST program parameters set to a score of 50 and a word length of -3. To obtain gapped alignments for comparison, the gapped BLAST described in Altschul et al., Nucleic Acids Res., 1997, 25:3389-402 can be used. Alternatively, PSI BLAST can be used to perform iterative searches to detect distant relatives between molecules (as described above). When using the BLAST program, the gapped BLAST program, and the PSI BLAST program, the default parameters for each program (e.g., XBLAST and NBLAST) can be used (see, for example, the National Center for Biotechnology Information (NCBI) at ncbi.nlm.nih.gov on the World Wide Web). Another non-restrictive example of a mathematical algorithm used for sequence comparison is the algorithm of Myers and Miller, 1988, CABIOS 4:11 17. Such an algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When using the ALIGN program to compare amino acid sequences, the PAM120 weight residue table, gap length penalty 12, and gap penalty 4 can be used.

[0152] The term “Subject” means, as used herein, “animal,” specifically “mammal,” for example, non-primates (e.g., mice, rats, cattle, horses, domestic cats, tigers and other megafelines, dogs, pigs, rabbits, goats, deer, sheep, polecats, gerbils, guinea pigs, hamsters, bats, and birds (e.g., chickens, turkeys, and ducks)) or primates (e.g., monkeys, baboons, chimpanzees, and humans). This term may be used interchangeably with the terms “patient” or “individual.” In some embodiments, the subject is a mammal diagnosed with a disease or disorder provided herein, for example, a human. In some embodiments, the subject is a mammal at risk of developing a disease or disorder provided herein, for example, a human. In some embodiments, the subject is a human.

[0153] The term “synthetic” as applied to nucleic acids, polypeptides, cells, or organisms refers to nucleic acids, polypeptides, cells, or organisms that cannot be directly isolated from their natural origin. In some embodiments, synthetic nucleic acids, polypeptides, cells, or organisms are substantially similar to their naturally occurring counterparts. In some embodiments, synthetic nucleic acids, polypeptides, cells, or organisms are modified or altered compared to their naturally occurring counterparts. In some embodiments, synthetic nucleic acids, polypeptides, cells, or organisms are made by functionally linking or combining various fragments of nucleic acids, polypeptides, cells, or organisms of origin. For example, synthetic polypeptides may include polypeptides of different origins that have been functionally linked.

[0154] The terms “treatment” and “to treat” refer, as used herein, to a pharmaceutical or other intervention plan for obtaining a beneficial or desired outcome in a recipient. Beneficial or desired outcomes include, but are not limited to, therapeutic and / or preventive benefits. A therapeutic benefit may refer to the disappearance or alleviation of the symptom or underlying disorder being treated. A therapeutic benefit may also be achieved by the disappearance or alleviation of one or more physiological symptoms associated with the underlying disorder, even if the subject still suffers from the underlying disorder, such that improvement is observed in the subject. Preventive effects include delaying, preventing, or eliminating the onset of a disease or condition; delaying or eliminating the onset of symptoms of a disease or condition; slowing, stopping, or reversing the progression of a disease or condition; or a combination thereof. For a preventive benefit, subjects at risk of developing a particular disease, or those reporting one or more physiological symptoms of a disease, may receive treatment even if they have not been diagnosed with the disease.

[0155] The term “vector,” as used herein, refers to a substance used to transport or introduce a nucleic acid sequence (e.g., a nucleic acid sequence encoding a synthetic cytokine receptor as described herein) into a host cell. Applicable vectors for use include, for example, plasmids, phage vectors, viral vectors, episomes, and artificial chromosomes. A vector may contain a sequence that directs autonomous replication within the cell, or it may contain a sequence sufficient to enable integration into host cell DNA. Furthermore, a vector may contain one or more selectable marker genes and appropriate expression regulatory sequences. Possible selectable marker genes may, for example, provide resistance to antibiotics or toxins, compensate for nutrient deficiencies, or supply essential nutrients that are not present in the culture medium. Expression regulatory sequences may include constitutive and inductive promoters, transcription enhancers, transcription termination factors, etc., which are well known in the art. When it is desired to co-express two or more nucleic acid molecules (e.g., the heavy and light chains of an antibody, or both the VH and VL of an antibody), both nucleic acid molecules may be inserted, for example, into a single expression vector, or into separate expression vectors. In single-vector expression, the coding nucleic acid may be functionally ligated to one common regulatory expression sequence or to different regulatory expression sequences. The introduction of the nucleic acid molecule into host cells can be confirmed using methods well known in the art. Such methods include, for example, nucleic acid analysis, such as Northern blotting or polymerase chain reaction (PCR) amplification of mRNA, immunoblotting for gene product expression, or other suitable analytical methods for testing the expression of the introduced nucleic acid sequence or the corresponding gene product. It is understood by those skilled in the art that nucleic acid molecules are expressed in sufficient quantities to produce the desired product, and it is also understood that expression levels can be optimized to obtain sufficient expression using methods well known in the art.

[0156] Common methods in molecular and cellular biochemistry are found in: Molecular Cloning: A Laboratory Manual, 3rd Ed. (Sambrook et al., Harbor Laboratory Press 2001); Short Protocols in Molecular Biology, 4th Ed. (Ausubel et al. eds., John Wiley & Sons 1999); Protein Methods (Bollag et al., John Wiley & Sons 1996); Nonviral Vectors for Gene Therapy (Wagner et al. eds., Academic Press 1999); Viral Vectors (Kaplift & Loewy eds., Academic Press 1995); Immunology Methods Manual (I. Lefkovits ed., Academic Press 1997); and Cell and Tissue Culture: Laboratory Procedures in Biotechnology (Doyle & Griffiths, John Wiley & Sons). These disclosures can be found in standard textbooks such as the one from 1998, and are incorporated herein by reference.

[0157] II. Synthetic cytokine receptors Synthetic cytokine receptors are provided herein. In some embodiments, the synthetic cytokine receptors are interleukin-9 receptor (IL-9R) signaling capable.

[0158] In some embodiments, synthetic cytokine receptors comprise an extracellular domain, a transmembrane domain, and an interleukin-9 receptor (IL-9R) intracellular domain. In some embodiments, the synthetic cytokine receptors provided herein can multimerize, typically as homodimers, even in the absence of external stimuli to the receptor, such as in the absence of ligand binding to the extracellular domain, thereby facilitating downstream signaling. Thus, in some embodiments, synthetic cytokine receptors are constitutively active. In some embodiments, the multimerization (e.g., dimerization) of synthetic cytokine receptors allows the receptor to be constitutively active by directing the polypeptide chain of the receptor so that JAK / STAT signaling molecules are recruited for subsequent activation. Typically, IL-9R signaling requires heterodimerization by a common γ-chain subunit, activation of JAK kinases (e.g., JAK1 and JAK3) for association with the receptor chain and its phosphorylation, and binding of its cytokines to the IL-9Rα chain to promote the subsequent phosphorylation and recruitment of other signaling molecules, e.g., STAT transcription factors, specifically STAT1, STAT3, and STAT5. In the provided embodiment, the synthetic receptor spontaneously forms homomultimers, e.g., homodimers, which constitutively activate JAK signaling, resulting in the activation of STAT1, STAT3, and / or STAT5, including the transposition of homodimers and heterodimers (e.g., STAT-1 / STAT-3) of STAT-1, STAT-3, or STAT-5 into the nucleus to regulate gene expression.

[0159] In some embodiments, modified cells (e.g., T cells) expressing the synthetic cytokine receptors provided herein, and in some cases also expressing chimeric antigen receptors (CARs), maintain their proliferative capacity even in the absence of extracellular domain ligands (e.g., in the absence of cytokines).

[0160] In some embodiments, synthetic cytokine receptors contain a multimer. In some embodiments, the multimer is a dimer.

[0161] In some aspects, synthetic cytokine receptors are polymerized to a higher order (e.g., trimers, tetramers, or more). In some aspects, synthetic cytokine receptors are polymerized as trimers, tetramers, pentamers, hexamers, heptamers, octamers, nonamers, or decamers. In some aspects, the polymer is a trimer. In some aspects, the polymer is a tetramer. In some aspects, the polymer is a pentamer. In some aspects, the polymer is a hexamer. In some aspects, the polymer is a heptamer. In some aspects, the polymer is an octamer. In some aspects, the polymer is a nonamer. In some aspects, the polymer is a decamer. In some aspects, the polymer is an elevenmer. In some aspects, the polymer is a dodecamer.

[0162] In some embodiments, the multimer contains an identical polypeptide chain comprising an extracellular domain, a transmembrane domain, and an IL-9R intracellular domain, respectively. In some embodiments, the multimer is a homomultimer. In some embodiments, the homomultimer is a homodimer. In some embodiments, the homomultimer is a homotrimer. In some embodiments, the homomultimer is a homotetramer. In some embodiments, the homomultimer is a homopentamer. In some embodiments, the homomultimer is a homohexamer. In some embodiments, the homomultimer is a homoheptamer. In some embodiments, the homomultimer is a homooctamer. In some embodiments, the homomultimer is a homonotamer. In some embodiments, the homomultimer is a homodecamer. In some embodiments, the homomultimer is a homodecamer. In some embodiments, the homomultimer is a homodecamer. In some embodiments, the homomultimer is a homodecamer. In some embodiments, the homomultimer is a homodecamer. In some embodiments, the homomultimer is a homodecamer. In some embodiments, the multimer is a heteromultimer.

[0163] In some embodiments, synthetic cytokine receptors are multimerized through an extracellular domain, a transmembrane domain, and / or an IL-9R intracellular domain. In some embodiments, synthetic cytokine receptors are multimerized through an extracellular domain. In some embodiments, synthetic cytokine receptors are multimerized through a transmembrane domain. In some embodiments, synthetic cytokine receptors are multimerized through an IL-9R intracellular domain. In some embodiments, synthetic cytokine receptors are multimerized through both an extracellular domain and a transmembrane domain. In some embodiments, synthetic cytokine receptors are multimerized through both a transmembrane domain and an IL-9R intracellular domain. In some embodiments, synthetic cytokine receptors are multimerized through both an extracellular domain and an IL-9R intracellular domain. In some embodiments, synthetic cytokine receptors are multimerized through an extracellular domain, a transmembrane domain, and an IL-9R intracellular domain.

[0164] In some embodiments, synthetic cytokine receptors are constitutively active cytokine receptors. In some embodiments, synthetic cytokine receptors are polymers of constitutively polymerized polypeptide chains. In some embodiments, constitutively polymerized synthetic cytokine receptors are constitutively active. In some embodiments, polymerized synthetic cytokine receptors are constitutively active because each polypeptide chain in the polymer enables polymerization without the need for external stimulation of the receptor. In some embodiments, constitutively polymerized synthetic cytokine receptors induce signal transduction via the STAT pathway.

[0165] In some embodiments, an advantage of the synthetic cytokine receptors provided herein is that the receptor is constitutively active even in the absence of a ligand. That is, downstream IL-9R signaling is constitutive even in the absence of ligand binding to the ECD of the synthetic cytokine receptor. However, ligand binding to the ECD of the synthetic cytokine receptors provided herein is not excluded. In some embodiments, the ECD of the synthetic cytokine receptors provided herein binds to a congener ligand. However, the constitutive signaling of the provided synthetic cytokine receptors is independent of ECD ligand binding. Therefore, in some embodiments, the synthetic cytokine receptors provided herein are constitutively active in the presence and absence of one or more ligands. In some embodiments, the synthetic cytokine receptors provided herein are constitutively active in the presence and absence of any ligand. In a specific embodiment, when the ECD of the synthetic cytokine receptors provided herein is derived from the CD34 receptor, the synthetic cytokine receptor is constitutively active in the presence and absence of the CD34 receptor ligand. In specific embodiments, when the ECD of the synthetic cytokine receptor provided herein is derived from the DAP12 receptor, the synthetic cytokine receptor is constitutively active in the presence and absence of the DAP12 receptor ligand. In specific embodiments, when the ECD of the synthetic cytokine receptor provided herein is derived from the GpA receptor, the synthetic cytokine receptor is constitutively active in the presence and absence of the GpA receptor ligand. In specific embodiments, when the ECD of the synthetic cytokine receptor provided herein is derived from the CD8 receptor, the synthetic cytokine receptor is constitutively active in the presence and absence of the CD8 receptor ligand. In specific embodiments, when the ECD of the synthetic cytokine receptor provided herein is derived from the Muc24 receptor, the synthetic cytokine receptor is constitutively active in the presence and absence of the Muc24 receptor ligand. In specific embodiments, the ECD does not substantially bind to the native ligand of the receptor from which the ECD is derived, for example, not specifically or with high affinity.In some embodiments, the native ligand of the receptor derived from ECD does not bind to the ECD of the synthetic cytokine receptor in a manner that can modulate intracellular signaling (e.g., STAT signaling) by the synthetic cytokine receptor. In specific embodiments, the synthetic cytokine receptor provided herein induces IL-9R signaling in the absence of IL-9. In some embodiments, the synthetic cytokine receptor provided herein does not bind to IL-9.

[0166] In some embodiments, a constitutively multimerized synthetic cytokine receptor is a constitutively active cytokine receptor that induces signaling through the STAT1, STAT3, or STAT5 pathway. In some embodiments, a constitutively multimerized synthetic cytokine receptor induces signaling through the STAT1 pathway. In some embodiments, a multimerized synthetic cytokine receptor induces signaling through the STAT3 pathway. In some embodiments, a constitutively multimerized synthetic cytokine receptor induces signaling through the STAT5 pathway. In some embodiments, a constitutively multimerized synthetic cytokine receptor simultaneously induces signaling through the STAT1, STAT3, and STAT5 pathways. In some embodiments, a constitutively multimerized synthetic cytokine receptor induces signaling through the STAT1, STAT3, and STAT5 pathways in any order. Signaling through STAT1, STAT3, and / or STAT5 can be determined by any method known in the art. In some embodiments, signaling through STAT1, STAT3, and / or STAT5 is determined by detecting phosphorylated STAT1, STAT3, and / or STAT5. For example, phosphorylation of STAT1, STAT3, and / or STAT5 can be detected by Western blotting using antibodies that specifically bind to phosphorylated STAT1, STAT3, and / or STAT5.

[0167] In some embodiments, synthetic cytokine receptors that multimerize to a higher order (e.g., trimers, tetramers, or more) induce stronger signaling compared to dimerized synthetic cytokine receptors. In some embodiments, signaling induced by multimerized synthetic cytokine receptors is at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% stronger than that induced by dimerized synthetic cytokine receptors. In some embodiments, signaling induced by multimerized synthetic cytokine receptors is at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, or at least about 9 times stronger than that induced by dimerized synthetic cytokine receptors. In some embodiments, signaling induced by trimer, tetramer, pentamer, hexamer, heptamer, octamer, nonamer, decamer, decamer, and / or dodecamer synthetic cytokine receptors is at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% more potent than that of dimerized synthetic cytokine receptors. In some embodiments, signaling induced by trimer, tetramer, pentamer, hexamer, heptamer, octamer, nonamer, decamer, decamer, and / or dodecamer synthetic cytokine receptors is at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, or at least about 9 times more potent than that of dimerized synthetic cytokine receptors.

[0168] In some embodiments, stronger signaling may be demonstrated by sustained or prolonged STAT activation. In some embodiments, STAT activation is indicated by detecting phosphorylated STAT. In some embodiments, phosphorylation of STAT1, STAT3, and / or STAT5 may be detected by Western blotting using antibodies that specifically bind to phosphorylated STAT1, STAT3, and / or STAT5. In some embodiments, signaling through STAT1, STAT3, and / or STAT5 may be determined by any method known in the art.

[0169] In some embodiments, the multimerization of synthetic cytokine receptors occurs through covalent or non-covalent interactions.

[0170] In some embodiments, the multimerization of synthetic cytokine receptors occurs through covalent interactions. In some embodiments, each synthetic cytokine receptor in the multimer is linked to another via one or more covalent bonds. In some embodiments, the covalent bond is a disulfide bond. In some embodiments, the disulfide bond is formed between cysteine ​​residues.

[0171] In some embodiments, synthetic cytokine receptors include a self-assembly domain that facilitates receptor multimerization. Self-assembly is the spontaneous association of a group of molecules into one or more supramolecular structures, driven by multiple non-covalent interactions. In some embodiments, multimerization of synthetic cytokine receptors occurs through non-covalent interactions. In some embodiments, non-covalent interactions are facilitated by amino acid sequence motifs. In some embodiments, amino acid sequence motifs facilitate dimerization or multimerization via α-helices. In some embodiments, amino acid sequence motifs facilitate dimerization or multimerization via leucine zippers.

[0172] References to amino acids, including references to specific sequences described as SEQ ID NOs used to describe domain structures (e.g., extracellular, transmembrane, or intracellular domains), are for illustrative purposes only and should not limit the scope of the embodiments provided. Descriptions of polypeptides and their domains are theoretically derived based on homology analysis and alignment with similar molecules. Therefore, precise locations may vary and are not necessarily the same for each protein. Thus, a particular domain may be several amino acids longer or shorter (e.g., 1, 2, 3, or 4 amino acids). Typically, transmembrane domains identified for polypeptides herein are identified according to criteria routinely used in the art for identifying the type of hydrophobic domain. While the precise boundaries of transmembrane domains may vary, such variation is likely to be less than approximately 5 amino acids at each end of the initially identified domain. In some embodiments, extracellular domains may be soluble (i.e., not membrane-bound) when not accompanied by transmembrane and cytoplasmic domains. The intracellular signaling domain is understood to be an IL-9R sequence or its variants or portions that exhibit the ability to induce intracellular signaling by recruiting, for example, JAK molecules and STAT molecules (e.g., STAT1, STAT3, and STAT5).

[0173] In some embodiments, the different domains of a synthetic cytokine receptor (i.e., the extracellular domain, the transmembrane domain, and the intracellular domain) are functionally linked. In some embodiments, the different elements are directly linked to each other. In some embodiments, the different elements are linked via a peptide linker, for example, a (GxS)n linker (where x can be an integer from 1 to 10 and n can be an integer from 2 to 20). Additional linkers are known to those skilled in the art.

[0174] A. Extracellular domain In some embodiments, the extracellular domain includes the extracellular domain of a naturally occurring protein. In some embodiments, the extracellular domain includes a fragment or truncated portion of the extracellular domain of a naturally occurring protein.

[0175] In some embodiments, the extracellular domain of the provided synthetic cytokine receptor allows the polypeptide chain to be constitutively active by enabling the polypeptide chain to multimerize (e.g., dimerize) so that the JAK / STAT molecule is recruited by the downstream intracellular domain of the polypeptide chain, even in the absence of external stimuli, for example, in the absence of ligand binding to the extracellular domain.

[0176] In some embodiments, the extracellular domain may or may not originate from the same native molecule as the functionally linked transmembrane domain. In some embodiments, the extracellular domain originates from the same molecule as the corresponding transmembrane domain. In some embodiments, the extracellular domain does not originate from the same molecule as the corresponding transmembrane domain.

[0177] In some embodiments, the extracellular domain contains one or more mutations compared to the corresponding wild-type protein. In some embodiments, the mutations may include substitutions, insertions, deletions, or combinations thereof.

[0178] In some embodiments, the extracellular domain interacts with one or more additional extracellular domains to promote the multimerization of synthetic cytokine receptors.

[0179] In some embodiments, the extracellular domain stabilizes synthetic cytokine receptors expressed on the cell surface. In some embodiments, the extracellular domain enhances downstream signaling induced by the IL-9R intracellular domain. In some embodiments, the extracellular domain confers a sink or ligand-trapping function, such as binding to and capturing one or more molecules harmful to cells expressing the synthetic cytokine receptor. In some embodiments, the ligand is immunosuppressive. Examples of harmful ligands include TGFβ, PD-L1, IL-4, IL-13, IL-8, and IL-10.

[0180] In some embodiments, the extracellular domains disclosed herein can bind to a ligand, but the signal of that ligand is not transmitted.

[0181] In some aspects, the size of the extracellular domain is at least about 30 amino acids. In some aspects, the size of the extracellular domain is up to about 300 amino acids in length. In some aspects, the size of the extracellular domain is about 30 to about 300 amino acids. In some aspects, the size of the extracellular domain is about 30 to about 50 amino acids, about 30 to about 70 amino acids, about 30 to about 90 amino acids, about 30 to about 110 amino acids, about 30 to about 130 amino acids, about 30 to about 150 amino acids, about 30 to about 170 amino acids, about 30 to about 190 amino acids, about 30 to about 210 amino acids, about 30 to about 250 amino acids, about 30 to about 300 amino acids, and about 50 amino acids. Acid ~ approximately 70 amino acids, approximately 50 amino acids ~ approximately 90 amino acids, approximately 50 amino acids ~ approximately 110 amino acids, approximately 50 amino acids ~ approximately 130 amino acids, approximately 50 amino acids ~ approximately 150 amino acids, approximately 50 amino acids ~ approximately 170 amino acids, approximately 50 amino acids ~ approximately 190 amino acids, approximately 50 amino acids ~ approximately 210 amino acids, approximately 50 amino acids ~ approximately 250 amino acids, approximately 50 amino acids ~ approximately 300 amino acids, approximately 70 amino acids ~ approximately 90 amino acids, approximately 70 amino acids ~ approximately 110 amino acids, approximately 70 amino acids ~ approximately 130 amino acids, approximately 70 amino acids Mino acids ~ approximately 150 amino acids, approximately 70 amino acids ~ approximately 170 amino acids, approximately 70 amino acids ~ approximately 190 amino acids, approximately 70 amino acids ~ approximately 210 amino acids, approximately 70 amino acids ~ approximately 250 amino acids, approximately 70 amino acids ~ approximately 300 amino acids, approximately 90 amino acids ~ approximately 110 amino acids, approximately 90 amino acids ~ approximately 130 amino acids, approximately 90 amino acids ~ approximately 150 amino acids, approximately 90 amino acids ~ approximately 170 amino acids, approximately 90 amino acids ~ approximately 190 amino acids, approximately 90 amino acids ~ approximately 210 amino acids, approximately 90 amino acids ~ approximately 250 amino acids Approximately 90 to 300 amino acids, approximately 110 to 130 amino acids, approximately 110 to 150 amino acids, approximately 110 to 170 amino acids, approximately 110 to 190 amino acids, approximately 110 to 210 amino acids, approximately 110 to 250 amino acids, approximately 110 to 300 amino acids, approximately 130 to 150 amino acids, approximately 130 to 170 amino acids, approximately 130 to 190 amino acids, approximately 130 to 210 amino acids,The amino acids range from approximately 130 to 250 amino acids, 130 to 300 amino acids, 150 to 170 amino acids, 150 to 190 amino acids, 150 to 210 amino acids, 150 to 250 amino acids, 150 to 300 amino acids, 170 to 190 amino acids, 170 to 210 amino acids, 170 to 250 amino acids, 170 to 300 amino acids, 190 to 210 amino acids, 190 to 250 amino acids, 190 to 300 amino acids, 210 to 250 amino acids, 210 to 300 amino acids, or 250 to 300 amino acids. In some embodiments, the size of the extracellular domain is approximately 30 amino acids, 50 amino acids, 70 amino acids, 90 amino acids, 110 amino acids, 130 amino acids, 150 amino acids, 170 amino acids, 190 amino acids, 210 amino acids, 250 amino acids, or 300 amino acids.

[0182] In some embodiments, the extracellular domain is approximately 10 to 260 amino acids long. In some embodiments, the extracellular domain is approximately 60 to 260 amino acids long. In some embodiments, the extracellular domain is approximately 69 amino acids long. In some embodiments, the extracellular domain is approximately 72 amino acids long. In some embodiments, the extracellular domain is approximately 139 amino acids long. In some embodiments, the extracellular domain is approximately 259 amino acids long.

[0183] In some embodiments, the extracellular domain includes a dimerization domain. In some embodiments, the dimerization domain includes a hinge region. In some embodiments, the extracellular domain facilitates disulfide bond dimerization. In some embodiments, disulfide bond dimerization is caused or facilitated by one or more cysteine ​​residues. In some embodiments, the extracellular domain includes 1 to 6 cysteine ​​residues. In some embodiments, the extracellular domain includes 1 cysteine ​​residue. In some embodiments, the extracellular domain includes 2 cysteine ​​residues. In some embodiments, the extracellular domain includes 3 cysteine ​​residues. In some embodiments, the extracellular domain includes 4 cysteine ​​residues. In some embodiments, the extracellular domain includes 5 cysteine ​​residues. In some embodiments, the extracellular domain includes 6 cysteine ​​residues. In some embodiments, disulfide bond dimerization forms 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor. In some embodiments, disulfide bond dimerization forms 1 disulfide crosslink between the polypeptide chains of the synthetic cytokine receptor. In some embodiments, disulfide dimerization forms two disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor. In some embodiments, disulfide dimerization forms three disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor. In some embodiments, the disulfide dimerization domain forms four disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

[0184] In some embodiments, the extracellular domain includes an extracellular domain derived from CD8, CD34, Muc24, DAP12, or glycophorin A (GpA). In some embodiments, the extracellular domain is derived from the extracellular domain of CD34. In some embodiments, the extracellular domain is derived from the extracellular domain of DAP12. In some embodiments, the extracellular domain is derived from the extracellular domain of glycophorin A. In some embodiments, the extracellular domain is derived from the extracellular domain of CD8. In some embodiments, the extracellular domain is derived from the extracellular domain of Muc24.

[0185] In some embodiments, the extracellular domain comprises the extracellular domain of CD8 or a truncated portion thereof. In some embodiments, the CD8 extracellular domain comprises at least one cysteine ​​residue.

[0186] In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:18. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:18.

[0187] In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 70% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 75% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 80% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 86% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 87% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 88% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 89% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 90% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 91% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 92% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 93% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 94% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 95% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 96% identical to that of SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least about 97% identical to that of SEQ ID NO:18.In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:18. In some embodiments, the CD8 extracellular domain contains the amino acid sequence shown in SEQ ID NO:18. In some embodiments, the CD8 extracellular domain consists of the amino acid sequence shown in SEQ ID NO:18.

[0188] In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to that of SEQ ID NO:19. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 70% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 75% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 80% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 85% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 86% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 87% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 88% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 89% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 90% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 91% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 92% identical to that of SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 93% identical to that of SEQ ID NO:19. In some embodiments, the extracellular domain of CD8 contains an amino acid sequence that is at least approximately 94% identical to that of SEQ ID NO:19. In some embodiments, the extracellular domain of CD8 contains an amino acid sequence that is at least approximately 95% identical to that of SEQ ID NO:19.In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:19. In some embodiments, the CD8 extracellular domain contains the amino acid sequence shown in SEQ ID NO:19. In some embodiments, the CD8 extracellular domain consists of the amino acid sequence shown in SEQ ID NO:19. In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:185. In some embodiments, the CD8 extracellular domain contains the amino acid sequence shown in SEQ ID NO:185. In some embodiments, the CD8 extracellular domain consists of the amino acid sequence shown in SEQ ID NO:185.

[0189] In some embodiments, the extracellular domain comprises the extracellular domain of CD34 or a truncated portion thereof. In some embodiments, the CD34 extracellular domain comprises at least one cysteine ​​residue. In some embodiments, the extracellular domain comprises an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:4. In some embodiments, the extracellular domain comprises the amino acid sequence of SEQ ID NO:4.

[0190] In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least about 70% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least about 75% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least about 80% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least about 86% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least about 87% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least about 88% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least about 89% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 90% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 91% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 92% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 93% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 94% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 95% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 96% identical to that of SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 97% identical to that of SEQ ID NO:4.In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:4. In some embodiments, the CD34 extracellular domain contains the amino acid sequence shown in SEQ ID NO:4. In some embodiments, the CD34 extracellular domain consists of the amino acid sequence shown in SEQ ID NO:4.

[0191] In some embodiments, the extracellular domain is the extracellular domain of Muc24 or a truncated portion thereof. In some embodiments, the Muc24 extracellular domain contains at least one cysteine ​​residue. In some embodiments, the extracellular domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:20. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:20.

[0192] In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 70% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 75% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 80% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 86% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 87% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 88% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 89% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 90% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 91% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 92% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 93% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 94% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 95% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 96% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:20.In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain contains the amino acid sequence shown in SEQ ID NO:20. In some embodiments, the Muc24 extracellular domain consists of the amino acid sequence shown in SEQ ID NO:20.

[0193] In some embodiments, the extracellular domain is the extracellular domain of DAP12 or a truncated portion thereof. In some embodiments, the DAP12 extracellular domain contains at least one cysteine ​​residue. In some embodiments, the extracellular domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:5. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:5.

[0194] In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 70% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 75% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 80% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 86% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 87% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 88% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 89% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 90% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 91% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 92% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 93% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 95% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 95% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 96% identical to that of SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least about 97% identical to that of SEQ ID NO:5.In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain contains the amino acid sequence shown in SEQ ID NO:5. In some embodiments, the DAP12 extracellular domain consists of the amino acid sequence shown in SEQ ID NO:5.

[0195] In some embodiments, the extracellular domain is the extracellular domain of GpA or a truncated portion thereof. In some embodiments, the extracellular domain of GpA contains at least one cysteine ​​residue.

[0196] In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of SEQ ID NO:16. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:16.

[0197] In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 70% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 75% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 80% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 86% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 87% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 88% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 89% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO:16.In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:16. In some embodiments, the GpA extracellular domain contains the amino acid sequence shown in SEQ ID NO:16. In some embodiments, the GpA extracellular domain consists of the amino acid sequence shown in SEQ ID NO:16.

[0198] In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to that of SEQ ID NO:17. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:17.

[0199] In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 70% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 75% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 80% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 86% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 87% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 88% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least about 89% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO:17.In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:17. In some embodiments, the GpA extracellular domain contains the amino acid sequence shown in SEQ ID NO:17. In some embodiments, the GpA extracellular domain consists of the amino acid sequence shown in SEQ ID NO:17.

[0200] In some embodiments, the extracellular domain is the active leucine zipper domain of the transcription factor. In some embodiments, the transcription factor is a basic leucine zipper (bZIP). Non-limiting examples of bZIP transcription factors that may be used herein include ATF, JUN, CREB, and Fos. In some embodiments, the extracellular domain is the leucine zipper domain of ATF. In some embodiments, the extracellular domain is the leucine zipper domain of JUN. In some embodiments, the extracellular domain is the leucine zipper domain of CREB. In some embodiments, the extracellular domain is the leucine zipper domain of Fos.

[0201] In some embodiments, the leucine zipper includes Leu-X6-Leu-X6-Leu-X6-Leu (SEQ ID NO: 72). In some embodiments, the leucine zipper is Includes TIFF2026525229000005.tif4128. In some embodiments, the leucine zipper is Includes TIFF2026525229000006.tif4128.

[0202] In some embodiments, the extracellular domain is the extracellular domain of the thrombopoietin receptor (TpoR) or a truncated portion thereof. In some embodiments, the extracellular domain is a truncated portion of TpoR. In some embodiments, the truncated TpoR extracellular domain contains the deletion of up to 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acid residues from the N-terminus of the wild-type TpoR extracellular domain shown in SEQ ID NO:178.

[0203] In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:178. In some embodiments, the TpoR extracellular domain contains the amino acid sequence shown in SEQ ID NO:178. In some embodiments, the TpoR extracellular domain consists of the amino acid sequence shown in SEQ ID NO:178. In some embodiments, the extracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:162. In some embodiments, the extracellular domain contains the amino acid sequence of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least about 70% identical to SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least about 75% identical to SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least about 80% identical to SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least about 86% identical to SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least about 87% identical to SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least about 88% identical to SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 89% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 90% identical to that of SEQ ID NO:162.In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 91% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 92% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 93% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 94% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 95% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 96% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 97% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 98% identical to that of SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:162. In some embodiments, the TpoR extracellular domain contains the amino acid sequence shown in SEQ ID NO:162. In some embodiments, the TpoR extracellular domain consists of the amino acid sequence shown in SEQ ID NO:162.

[0204] In some embodiments, the extracellular domains provided herein may include any of the extracellular domains shown in Table 1. As shown, the exact location or residues corresponding to a given domain may vary, for example, depending on the method used to identify or classify that domain. In some cases, the N-terminal and / or C-terminal amino acids adjacent to a given extracellular domain may be included in the sequence of the synthetic cytokine receptor, insofar as the resulting synthetic cytokine receptor can constitutively multimerize (e.g., dimerize) to result in constitutive activation of downstream signaling molecules. It should therefore be understood that the examples of SEQ ID NOs in Table 1 should not be interpreted restrictively. For example, a particular extracellular domain may be several amino acids longer or shorter than the amino acid sequence shown in its respective SEQ ID NO, for example, 1 to 10, for example, 1, 2, 3, 4, 5, 6, or 7 amino acids longer or shorter. Variants of such SEQ ID NO (e.g., sequences exhibiting at least about 90% identity, at least about 91% identity, at least about 92% identity, at least about 93% identity, at least about 94% identity, at least about 95% identity, e.g., at least about 96% identity, 97% identity, 98% identity, or 99% identity to any of SEQ ID NO: 4, 5, 16, 17, 18, 19, 20, 73, 74, 162, 178, or 185) are also provided, and synthetic cytokine receptors resulting from containing such variant extracellular domains can constitutively multimerize (e.g., dimerize) to result in constitutive activation of downstream signaling molecules.

[0205] (Table 1) Exemplary extracellular domains TIFF2026525229000007.tif50128

[0206] B. transmembrane domain In some embodiments, the transmembrane domain of the synthetic cytokine receptor includes the transmembrane domain of a naturally occurring protein. In some embodiments, the transmembrane domain includes a fragment or truncated portion of the transmembrane domain of a naturally occurring protein.

[0207] In some embodiments, the transmembrane domain of the provided synthetic cytokine receptor enables the synthetic cytokine receptor to be constitutively active by allowing the polypeptide chain to multimerize (e.g., dimerize) so that the JAK / STAT molecule is recruited by the downstream intracellular domain of the polypeptide chain, even in the absence of external stimuli, for example, in the absence of ligand binding to the extracellular domain.

[0208] In some embodiments, the transmembrane domain may or may not originate from the same native molecule as the functionally linked extracellular domain. In specific embodiments, the transmembrane domain does not originate from the same native molecule as the functionally linked extracellular domain.

[0209] In some embodiments, the transmembrane domain contains one or more mutations compared to the transmembrane domain of the corresponding wild-type protein. In some embodiments, one or more mutations promote, induce, or facilitate multimerization of the transmembrane domain compared to the corresponding wild-type protein. In some embodiments, the transmembrane domain contains one or more mutations that alter the arrangement of the downstream intracellular domain compared to the wild-type protein. In some embodiments, the transmembrane domain contains one or more mutations that induce or facilitate multimerization of the transmembrane domain and alter the arrangement of the downstream intracellular domain. Multimerization and / or altered arrangement cause the downstream IL-9R intracellular domains to be signal-signal-readably directed toward each other.

[0210] Mutations can be substitutions, insertions, deletions, or combinations thereof. In some embodiments, one or more mutations include at least one cysteine. In some embodiments, one or more mutations include at least one proline. Methods for determining whether a particular mutation induces downstream signaling are known to those skilled in the art (e.g., assays for phosphorylation of STAT1, STAT3, or STAT5 after proliferation of receptor-possessing cells tested in the absence of growth factors). In some embodiments, one or more mutations in the transmembrane domain are the introduction of at least one cysteine ​​residue. In some embodiments, the introduction of at least one cysteine ​​residue, for example by amino acid substitution, induces disulfide bond formation in the transmembrane domain. In some embodiments, one or more mutations do not include the introduction of cysteine. For example, variant transmembrane domains that are constitutively active and signal transduction-enabled may be utilized for mutations that alter the conformation of the transmembrane domain compared to the native version of the transmembrane domain, thereby enabling signal transduction, even if they do not have cysteine ​​insertions or amino acid substitutions (e.g., for disulfide bonds with adjacent polypeptide chains). For example, an insertion or substitution of an amino acid such as proline may cause a "kink" that bends the transmembrane domain, which may induce signal transduction (see, e.g., Shochat et al, 2011, J Exp Med. 2011 May 9;208(5):901-8; Zenatti et al, 2011, Nat Genet. 2011 Sep 4;43(10):932-9). In some embodiments, one or more mutations in the transmembrane domain are the introduction of proline residues. In some embodiments, the mutation is or includes one or more cysteine ​​and / or one or more proline insertions or amino acid substitutions. In some embodiments, the mutation is caused by the introduction (e.g., insertion or amino acid substitution) of a trimer peptide consisting of cysteine, proline, cystine, and another amino acid other than proline (e.g., threonine) into the transmembrane domain.In some embodiments, trimer peptides confer disulfide bond formation between the -SH (thiol) groups of cysteine ​​residues of two molecules, enabling their homodimer formation (in specific embodiments, proline immediately following cysteine ​​helps to bend the homodimer in the correct direction). In some embodiments, the trimer peptide is a trimer peptide of cysteine, proline, and threonine, in some cases a CPT insertion, or in some cases a TCP insertion.

[0211] In some embodiments, the size of the transmembrane domain is at least about 15 amino acids. In some embodiments, the size of the transmembrane domain is about 15 to about 45 amino acids. In some embodiments, the size of the transmembrane domain is up to about 45 amino acids. In some aspects, the transmembrane domain size is approximately 15 to 20 amino acids, 15 to 25 amino acids, 15 to 30 amino acids, 15 to 35 amino acids, 15 to 40 amino acids, 15 to 45 amino acids, 20 to 25 amino acids, 20 to 30 amino acids, 20 to 35 amino acids, 20 to 40 amino acids, 20 to 45 amino acids, 25 to 30 amino acids, 25 to 35 amino acids, 25 to 40 amino acids, 25 to 45 amino acids, 30 to 35 amino acids, 30 to 40 amino acids, 30 to 45 amino acids, 35 to 40 amino acids, 35 to 45 amino acids, or 40 to 45 amino acids. In some embodiments, the size of the transmembrane domain is approximately 15 amino acids, approximately 20 amino acids, approximately 25 amino acids, approximately 30 amino acids, approximately 35 amino acids, approximately 40 amino acids, or approximately 45 amino acids.

[0212] In some embodiments, the transmembrane domain is approximately 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 amino acid long. In some embodiments, the transmembrane domain is 21 amino acid long. In some embodiments, the transmembrane domain is 22 amino acid long. In some embodiments, the transmembrane domain is 23 amino acid long. In some embodiments, the transmembrane domain is 24 amino acid long. In some embodiments, the transmembrane domain is 25 amino acid long. In some embodiments, the transmembrane domain is 26 amino acid long. In some embodiments, the transmembrane domain is 27 amino acid long. In some embodiments, the transmembrane domain is 28 amino acid long. In some embodiments, the transmembrane domain is 29 amino acid long. In some embodiments, the transmembrane domain is 30 amino acid long. In some embodiments, the transmembrane domain is 31 amino acid long. In some embodiments, the transmembrane domain is 32 amino acid long. In some embodiments, the transmembrane domain is 33 amino acid long.

[0213] In some embodiments, the transmembrane domain promotes dimerization.

[0214] In some embodiments, the transmembrane domain is derived from the transmembrane domain of the interleukin-7 receptor. In some embodiments, the transmembrane domain is derived from the transmembrane domain of glycophorin A. In some embodiments, the transmembrane domain is derived from the transmembrane domain of Muc24.

[0215] In some embodiments, the transmembrane domain comprises a transmembrane domain derived from glycophorin A (GpA), carnitine palmitoyltransferase 1 (CPT1), tumor necrosis factor receptor (TNFR), or Muc24. In some embodiments, the transmembrane domain is a variant or truncated form of the transmembrane domain derived from glycophorin A (GpA), carnitine palmitoyltransferase 1 (CPT1), tumor necrosis factor receptor (TNFR), or Muc24.

[0216] In some embodiments, the transmembrane domain promotes α-helix dimerization. In some embodiments, α-helix oligomerization, e.g., dimerization or trimerization, is driven by a sequence motif, which is a simple, recognizable amino acid sequence that promotes lateral interactions. In some embodiments, the transmembrane domain contains a proline-rich motif ΦPXΦ (SEQ ID NO: 75) (where Φ represents a hydrophobic residue, P is proline, and X is any amino acid, typically a nonpolar residue other than proline and glycine). In some embodiments, the transmembrane domain contains ΦTXXAΦ (where Φ represents a hydrophobic residue, T is threonine, X is any amino acid, typically a nonpolar residue other than proline and glycine, and A is alanine). In some embodiments, the transmembrane domain contains a GXXXG motif (SEQ ID NO: 68). In some embodiments, the transmembrane domain contains a GXXXA motif (SEQ ID NO: 69). In some embodiments, the transmembrane domain contains an AXXXA motif (SEQ ID NO: 77). In some embodiments, the transmembrane domain contains an AXXXS motif (SEQ ID NO: 78).

[0217] In some embodiments, the transmembrane domain includes motif GXXXG (SEQ ID NO:68). In some embodiments, the transmembrane domain is motif Includes TIFF2026525229000008.tif4128. In some embodiments, the transmembrane domain is motif GXXXG (SEQ ID NO:68) and motif Includes TIFF2026525229000009.tif4128.

[0218] In some embodiments, the transmembrane domain is a transmembrane domain derived from glycophorin A (GpA) or a variant thereof. In some embodiments, the transmembrane domain comprises a transmembrane domain derived from the transmembrane domain of glycophorin A (GpA). In some embodiments, the transmembrane domain is a variant of the transmembrane domain of glycophorin A (GpA). In some embodiments, the variant comprises one or more mutations compared to the wild-type GpA transmembrane domain, e.g., 1, 2, 3, 4, 5, or 6 mutations (e.g., amino acid substitutions). In some embodiments, one or more mutations promote α-helix dimerization. In some embodiments, the GpA transmembrane domain or variant GpA transmembrane domain comprises the motif GXXXG (SEQ ID NO: 68). In some embodiments, the GpA transmembrane domain or variant GpA transmembrane domain is motif Includes TIFF2026525229000010.tif4128. In some embodiments, the GpA transmembrane domain or variant GpA transmembrane domain includes motif GXXXG (SEQ ID NO:68) and motif Includes TIFF2026525229000011.tif4128.

[0219] In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:22. In some embodiments, the transmembrane domain contains the amino acid sequence of SEQ ID NO:22.

[0220] In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 70% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 75% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 80% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 86% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 87% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 88% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 89% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 90% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 91% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 92% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 93% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 94% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 95% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 96% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:22.In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:22. In some embodiments, the GpA transmembrane domain contains the amino acid sequence shown in SEQ ID NO:22. In some embodiments, the GpA transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:22.

[0221] In some embodiments, the transmembrane domain is a variant GpA transmembrane domain. In some embodiments, the transmembrane domain contains an amino acid sequence exhibiting at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence identical to SEQ ID NO:43 by at least about 70%. In some embodiments, the transmembrane domain contains an amino acid sequence identical to SEQ ID NO:43 by at least about 75%. In some embodiments, the transmembrane domain contains an amino acid sequence identical to SEQ ID NO:43 by at least about 80%. In some embodiments, the transmembrane domain contains an amino acid sequence identical to SEQ ID NO:43 by at least about 85%. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 86% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 87% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 88% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 89% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:43.In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 96% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:43. In some embodiments, the transmembrane domain contains the amino acid sequence shown in SEQ ID NO:43. In some embodiments, the transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:43.

[0222] In some embodiments, the transmembrane domain includes motifs GXXXG (SEQ ID NO: 68) and GXXXA (SEQ ID NO: 69).

[0223] In some embodiments, the transmembrane domain comprises a transmembrane domain derived from carnitine palmitoyltransferase 1 (CPT1). In some embodiments, the CPT1 transmembrane domain, or a variant of the CPT1 transmembrane domain, comprises motifs GXXXG (SEQ ID NO: 68) and GXXXA (SEQ ID NO: 69).

[0224] In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence exhibiting at least approximately 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence identical to SEQ ID NO:45 by at least approximately 70%. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence identical to SEQ ID NO:45 by at least approximately 75%. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence identical to SEQ ID NO:45 by at least approximately 80%. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence identical to SEQ ID NO:45 by at least approximately 85%. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 86% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 87% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 88% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 89% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:45.In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least about 96% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain contains the amino acid sequence shown in SEQ ID NO:45. In some embodiments, the CPT1 transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:45.

[0225] In some embodiments, the transmembrane domain promotes disulfide dimerization. In some embodiments, the transmembrane domain contains 1 to 6 cysteine ​​residues. In some embodiments, the transmembrane domain contains 1 cysteine ​​residue. In some embodiments, the transmembrane domain contains 2 cysteine ​​residues. In some embodiments, the transmembrane domain contains 3 cysteine ​​residues. In some embodiments, the transmembrane domain contains 4 cysteine ​​residues. In some embodiments, the transmembrane domain contains 5 cysteine ​​residues. In some embodiments, the transmembrane domain contains 6 cysteine ​​residues. In some embodiments, disulfide dimerization includes disulfide crosslinks. In some embodiments, disulfide dimerization includes 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor. In some embodiments, disulfide dimerization forms 1 disulfide crosslink between the polypeptide chains of the synthetic cytokine receptor. In some embodiments, disulfide dimerization forms 2 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor. In some embodiments, disulfide dimerization forms three disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor. In some embodiments, the disulfide dimerization domain forms four disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

[0226] In some embodiments, the transmembrane domain comprises a transmembrane domain derived from the interleukin 7 receptor (IL-7R). In some embodiments, the IL-7R transmembrane domain is a variant transmembrane domain. In some embodiments, the variant transmembrane domain comprises one or more mutations compared to the wild-type IL-7R transmembrane domain. In some embodiments, one or more mutations promote homodimerization of the synthetic cytokine receptor. In some embodiments, one or more mutations promote disulfide bond dimerization. In some embodiments, one or more mutations introduce at least one cysteine into the transmembrane domain. In some embodiments, one or more mutations introduce at least one proline into the transmembrane domain. In some embodiments, the mutation is or comprises an insertion or amino acid substitution of one or more cysteines and / or one or more prolines. In some embodiments, the mutation is by introduction (e.g., insertion or amino acid substitution) of a trimeric peptide of cysteine, proline, and another amino acid other than cystine and proline (e.g., threonine) into the IL-7R transmembrane domain. In some embodiments, the trimeric peptide is a trimeric peptide of cysteine, proline, and threonine, e.g., in some cases, CPT insertion or TCP insertion. In some embodiments, the transmembrane domain is the wild-type IL-7R transmembrane domain a transmembrane domain or portion thereof derived from IL-7R, or a contiguous sequence thereof of at least 21, 22, 23, 24, 25, 26, 27, or 28 amino acids, comprising one or more mutations compared to TIFF2026525229000012.tif4128. In some embodiments, the transmembrane domain may contain 1, 2, 3, or 4 additional amino acids at the N-terminus or C-terminus compared to the sequence shown in SEQ ID NO:71. In some embodiments, the transmembrane domain is the wild-type IL-7R transmembrane domain The variant transmembrane domain or portion thereof derived from IL-7R, containing one or more mutations compared to TIFF2026525229000013.tif4128. In some embodiments, the variant transmembrane domain containing one or more mutations compared to the wild-type IL-7R transmembrane domain is not 100% identical to SEQ ID NO:71, but has a sequence that is at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, or 95% identical to SEQ ID NO:71.

[0227] In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:6. In some embodiments, the transmembrane domain contains the amino acid sequence of SEQ ID NO:6.

[0228] In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 70% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 75% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 80% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 86% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 87% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 88% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 89% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 90% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 91% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 92% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 93% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 94% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 95% identical to that of SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:6.In some embodiments, the variant IL-7R transmembrane domain comprises an amino acid sequence that is at least about 97% identical to SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain comprises an amino acid sequence that is at least about 98% identical to SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain comprises an amino acid sequence that is at least about 99% identical to SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain comprises the amino acid sequence set forth in SEQ ID NO:6. In some embodiments, the variant IL-7R transmembrane domain consists of the amino acid sequence set forth in SEQ ID NO:6.

[0229] In some embodiments, the transmembrane domain comprises an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:7. In some embodiments, the transmembrane domain comprises the amino acid sequence of SEQ ID NO:7.

[0230] In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 70% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 75% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 80% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 86% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 87% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 88% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 89% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 90% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 91% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 92% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 93% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 94% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 95% identical to that of SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:7.In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain contains the amino acid sequence shown in SEQ ID NO:7. In some embodiments, the variant IL-7R transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:7.

[0231] In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:21. In some embodiments, the transmembrane domain contains the amino acid sequence of SEQ ID NO:21.

[0232] In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 70% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 75% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 80% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 86% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 87% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 88% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 89% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 90% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 91% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 92% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 93% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 94% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 95% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:21.In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain contains the amino acid sequence shown in SEQ ID NO:21. In some embodiments, the variant IL-7R transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:21.

[0233] In some embodiments, the transmembrane domain is a transmembrane domain derived from a TNF receptor (TNFR) that naturally contains a transmembrane domain helix capable of oligomerization, e.g., dimerization or trimerization. In some embodiments, the transmembrane domain derived from TNFR is a transmembrane domain derived from TACI, cell death receptor 5 (DR5), p75NTR, Fas, TNFR1, TNFR2, or OX40. In some embodiments, by including such a transmembrane domain in the provided synthetic cytokine receptor, constitutive clustering of intracellular domains that recruit and activate downstream signaling proteins, e.g., JAK and STAT, becomes possible. In some embodiments, the transmembrane domain contains motifs that support oligomerization, e.g., dimerization and / or trimerization (see, for example, Zhao et al., 2020, Frontiers in Cell and Developmental Biology, 8.569684.10.3389 / fcell.2020.569684, incorporated herein by reference). In some embodiments, the transmembrane domain derived from TNFR contains a proline-rich motif ΦPXΦ (SEQ ID NO: 75) (where Φ represents a hydrophobic residue, P is proline, and X is any amino acid, typically a nonpolar residue other than proline and glycine). In some embodiments, the transmembrane domain derived from TNFR contains ΦTXXAΦ (where Φ represents a hydrophobic residue, T is threonine, X is any amino acid, typically a nonpolar residue other than proline and glycine, and A is alanine). In some embodiments, the transmembrane domain derived from TNFR contains a GXXXG motif (SEQ ID NO: 68). In some embodiments, the transmembrane domain derived from TNFR contains a GXXXA motif (SEQ ID NO: 69). In some embodiments, the transmembrane domain derived from TNFR contains an AXXXA motif (SEQ ID NO: 77). In some embodiments, the transmembrane domain derived from TNFR contains an AXXXS motif (SEQ ID NO: 78).

[0234] In some embodiments, the transmembrane domain includes a transmembrane domain derived from DR5. In some embodiments, the DR5 transmembrane domain includes an amino acid sequence exhibiting at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain includes an amino acid sequence identical to SEQ ID NO:46 by at least about 70%. In some embodiments, the DR5 transmembrane domain includes an amino acid sequence identical to SEQ ID NO:46 by at least about 75%. In some embodiments, the DR5 transmembrane domain includes an amino acid sequence identical to SEQ ID NO:46 by at least about 80%. In some embodiments, the DR5 transmembrane domain includes an amino acid sequence identical to SEQ ID NO:46 by at least about 85%. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 86% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 87% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 88% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 89% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:46.In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least about 95% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least about 96% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain contains the amino acid sequence shown in SEQ ID NO:46. In some embodiments, the DR5 transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:46. In some embodiments, any of such DR5 transmembrane domains contains motif ΦTXXAΦ (SEQ ID NO:76) and / or motif GXXXG (SEQ ID NO:68). In some embodiments, any of the DR5 transmembrane domains comprises the motif ΦTXXAΦ (SEQ ID NO: 76). In some embodiments, any of the DR5 transmembrane domains comprises the motif GXXXG (SEQ ID NO: 68). In some embodiments, any of the DR5 transmembrane domains comprises the motif ΦTXXAΦ (SEQ ID NO: 76) and the motif GXXXG (SEQ ID NO: 68).

[0235] In some embodiments, the transmembrane domain includes a transmembrane domain derived from TACI. In some embodiments, the TACI transmembrane domain includes an amino acid sequence exhibiting at least about 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:44. In some embodiments, the TACI transmembrane domain includes an amino acid sequence identical to SEQ ID NO:44 by at least about 70%. In some embodiments, the TACI transmembrane domain includes an amino acid sequence identical to SEQ ID NO:44 by at least about 75%. In some embodiments, the TACI transmembrane domain includes an amino acid sequence identical to SEQ ID NO:44 by at least about 80%. In some embodiments, the TACI transmembrane domain includes an amino acid sequence identical to SEQ ID NO:44 by at least about 85%. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 86% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 87% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 88% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 89% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:44.In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least about 95% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least about 96% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:44. In some embodiments, the TACI transmembrane domain contains the amino acid sequence shown in SEQ ID NO:44. In some embodiments, the TACI transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:44. In some embodiments, any of such TACI transmembrane domains contains motif AXXXS (SEQ ID NO:78).

[0236] In some embodiments, the transmembrane domain includes a transmembrane domain derived from Muc24.

[0237] In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:23. In some embodiments, the transmembrane domain contains the amino acid sequence of SEQ ID NO:23.

[0238] In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 70% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 75% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 80% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 86% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 87% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 88% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 89% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 90% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 91% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 92% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 93% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 94% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 95% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 96% identical to that of SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 97% identical to that of SEQ ID NO:23.In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain contains the amino acid sequence shown in SEQ ID NO:23. In some embodiments, the Muc24 transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:23.

[0239] In some embodiments, the transmembrane domain includes a transmembrane domain derived from the thrombopoietin receptor (TpoR). In some embodiments, the TpoR transmembrane domain is a variant transmembrane domain. In some embodiments, the variant transmembrane domain is a wild-type TpoR transmembrane domain Compared to TIFF2026525229000014.tif4128, it contains one or more mutations. In some embodiments, one or more mutations promote homodimerization of the synthetic cytokine receptor. In some embodiments, one or more mutations promote α-helix dimerization. In some embodiments, compared to the wild-type TpoR transmembrane domain, the variant transmembrane domain containing one or more mutations is not 100% identical to SEQ ID NO:169, but has a sequence that is at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:169. In some embodiments, one or more mutations replace at least one amino acid residue in the wild-type transmembrane domain with a leucine residue. In some embodiments, one or more mutations replace at least one amino acid residue in the wild-type transmembrane domain with an asparagine residue. In some embodiments, one or more mutations replace one or more amino acid residues in the wild-type transmembrane domain with a leucine residue and an asparagine residue. In some embodiments, the TpoR TMD contains the amino acid substitution H499L relative to wild-type TpoR (SEQ ID NO: 168) or H8L relative to the wild-type TpoR transmembrane domain (SEQ ID NO: 169). In some embodiments, the TpoR TMD contains the amino acid substitution S505N relative to wild-type TpoR or S14N relative to the wild-type TpoR transmembrane domain. In some embodiments, TpoR TMDs contain amino acid substitutions of H499L and S505N relative to wild-type TpoR, or H8L and S14N relative to the wild-type TpoR transmembrane domain.

[0240] In some embodiments, the transmembrane domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:169. In some embodiments, the transmembrane domain contains the amino acid sequence of SEQ ID NO:169.

[0241] In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 70% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 75% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 80% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 86% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 87% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 88% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 89% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 90% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 91% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 92% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 93% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 94% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 95% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 96% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:169.In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain contains the amino acid sequence shown in SEQ ID NO:169. In some embodiments, the TpoR transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:169.

[0242] In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:163. In some embodiments, the transmembrane domain contains the amino acid sequence of SEQ ID NO:163.

[0243] In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 70% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 75% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 80% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 86% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 87% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 88% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 89% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 90% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 91% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 92% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 93% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 94% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 95% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 96% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 97% identical to SEQ ID NO:163.In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 98% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains an amino acid sequence that is at least about 99% identical to SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain contains the amino acid sequence shown in SEQ ID NO:163. In some embodiments, the TpoR transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:163.

[0244] In some embodiments, the transmembrane domain includes a transmembrane domain derived from wild-type IL-9R. In some embodiments, the transmembrane domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:125. In some embodiments, the wild-type IL-9R transmembrane domain includes the amino acid sequence shown in SEQ ID NO:125. In some embodiments, the wild-type IL-9R transmembrane domain consists of the amino acid sequence shown in SEQ ID NO:125.

[0245] In some embodiments, the transmembrane domains provided herein may comprise any of the transmembrane domains shown in Table 2. As shown, the exact location or residues corresponding to a given domain may vary, for example, depending on the method used to identify or classify that domain. In some cases, the N-terminal and / or C-terminal amino acids adjacent to a given transmembrane domain may be included in the sequence of the synthetic cytokine receptor, insofar as the resulting receptor can constitutively multimerize (e.g., dimerize) to result in constitutive activation of downstream signaling molecules. It should therefore be understood that the examples of SEQ ID NOs in Table 2 should not be interpreted restrictively. For example, a particular transmembrane domain may be several amino acids longer or shorter than the amino acid sequence shown in its respective SEQ ID NO, for example, 1 to 10, for example, 1, 2, 3, 4, 5, 6, or 7 amino acids longer or shorter. Variants of such SEQ ID NO (e.g., sequences exhibiting at least about 90% identity, at least about 91% identity, at least about 92% identity, at least about 93% identity, at least about 94% identity, at least about 95% identity, e.g., at least about 96% identity, 97% identity, 98% identity, or 99% identity to any of SEQ ID NO: 6, 7, 21, 22, 23, 43, 44, 45, 46, 125, 163, or 169) are also provided, and synthetic cytokine receptors resulting from containing such variant transmembrane domains can constitutively multimerize (e.g., dimerize) to result in constitutive activation of downstream signaling molecules.

[0246] (Table 2) Exemplary transmembrane domains TIFF2026525229000015.tif57128

[0247] C. intracellular domain Aspects provided relate to synthetic cytokine receptors comprising an extracellular domain (e.g., any of those described in Section II.A), a transmembrane domain (e.g., any of those described in Section II.B), and an intracellular domain capable of interleukin 9 receptor (IL-9R) signaling. In some aspects, the intracellular domain comprises an IL-9R intracellular domain or a variant thereof. In some aspects, the intracellular domain comprises a chimeric JAK / STAT fusion domain.

[0248] 1. IL-9R intracellular domain or a variant thereof In some aspects, the intracellular domain comprises the intracellular domain of a naturally occurring IL-9R (e.g., as shown in SEQ ID NO:8). In some aspects, the intracellular domain comprises one or more mutations (e.g., one or more mutations compared to the sequence shown in SEQ ID NO:8) compared to a naturally occurring IL-9R. The one or more mutations can be substitutions, insertions, deletions, or combinations thereof. In some aspects, the one or more mutations promote downstream signaling. Methods for determining whether a particular mutation induces downstream signaling are known to those of skill in the art (e.g., assays for phosphorylation of STAT1, STAT3, or STAT5 after proliferation of cells bearing the receptor tested in the absence of growth factor).

[0249] In some embodiments, the intracellular domain is at least about 150 amino acids long. In some embodiments, the intracellular domain is up to about 250 amino acids long. In some embodiments, the intracellular domain is about 150 to about 250 amino acids long. In some embodiments, the intracellular domain is about 169, 220, 223, or 230 amino acids long. In some embodiments, the intracellular domain is about 169 amino acids long. In some embodiments, the intracellular domain is about 220 amino acids long. In some embodiments, the intracellular domain is about 223 amino acids long. In some embodiments, the intracellular domain is about 230 amino acids long. In some embodiments, the intracellular domain is 169 amino acids long. In some embodiments, the intracellular domain is 220 amino acids long. In some embodiments, the intracellular domain is 223 amino acids long. In some embodiments, the intracellular domain is 230 amino acids long.

[0250] In some embodiments, the IL-9R intracellular domain is either the wild-type IL-9R intracellular domain or a variant thereof containing one or more mutations compared to the wild-type IL-9R intracellular domain shown in SEQ ID NO:8. In some embodiments, the one or more mutations include one or more amino acid insertions, deletions, and / or substitutions. In some embodiments, the one or more mutations facilitate signaling through the STAT1, STAT3, and / or STAT5 pathways.

[0251] In some embodiments, the IL-9R intracellular domain or a variant thereof includes a box 1 motif and / or a box 2 motif.

[0252] In some embodiments, the IL-9R intracellular domain or its variants include a box-1 motif defined as having proline, an arbitrary amino acid residue, and a hydrophobic sequence preceding the proline. In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, contains a proline-rich box-1 motif that enables the binding of the JAK molecule. In some embodiments, the box-1 motif is FYQNVPSPA (corresponding to positions 10-18 of the sequence shown in SEQ ID NO:79; SEQ ID NO:8).

[0253] In some embodiments, the IL-9R intracellular domain or its variants include a box-2 motif, defined as having a positively charged amino acid following a cluster of hydrophobic amino acids. In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, contains a box-2 motif that enables the binding of STAT molecules.

[0254] In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, contains a conserved tyrosine residue at position 116 of the sequence shown in SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, further contains conserved proline at position 118 and conserved glutamine at position 119, respectively, relative to the positions shown in SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, contains a conserved glutamine residue at position 118 of the sequence shown in SEQ ID NO:8.

[0255] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence of SEQ ID NO:8.

[0256] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 75% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 80% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 85% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 86% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 87% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 88% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 89% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 90% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 91% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 92% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 93% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 94% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 95% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 96% identical to that of SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 97% identical to that of SEQ ID NO:8.In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:8. In some embodiments, the IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:8.

[0257] In some embodiments, the IL-9R intracellular domain contains a deletion of a sequence of amino acids compared to the wild-type IL-9R intracellular domain. In some embodiments, the mutation is a deletion, and the variant is a deletion-type IL-9R intracellular signaling domain that lacks one or more regions of the wild-type IL-9R intracellular signaling domain (e.g., one or more regions of the sequence shown in SEQ ID NO:8). In some embodiments, IL-9R lacks up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 sequence of amino acids present in the wild-type IL-9R intracellular domain (e.g., SEQ ID NO:8). In some embodiments, IL-9R lacks a region of 10 sequence of amino acids relative to the wild-type IL-9R intracellular domain. In some embodiments, the deleted region is a sequence not required for the functional activity of the IL-9R intracellular domain, and therefore, signaling mediated by IL-9R is preserved.

[0258] In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, contains a deletion of the amino acid NGNFQTWMGA (corresponding to the deletion of amino acids 29-38 relative to the position indicated by SEQ ID NO:82;SEQ ID NO:8). In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, contains a deletion of the amino acid HGAGVLLSQD (corresponding to the deletion of amino acids 39-48 relative to the position indicated by SEQ ID NO:81;SEQ ID NO:8). In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, contains a deletion of the amino acid TCGPARPWKS (corresponding to the deletion of amino acids 69-78 relative to the position indicated by SEQ ID NO:83;SEQ ID NO:8). In some embodiments, the IL-9R intracellular domain, including the variant IL-9R intracellular domain, contains a deletion of the amino acid ALGCYGGWHL (corresponding to a deletion of amino acids 154-163 relative to the position indicated by SEQ ID NO:80;SEQ ID NO:8). In some embodiments, one or more further mutations may be present in any of these truncated IL-9R intracellular domains.

[0259] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence exhibiting at least approximately 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:51 by at least approximately 70%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:51 by at least approximately 75%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:51 by at least approximately 80%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:51 by at least approximately 85%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 86% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 87% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 88% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 89% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:51.In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO: 51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO: 51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO: 51. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO: 51. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO: 51. In some embodiments, the IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 51.

[0260] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence exhibiting at least approximately 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with respect to SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:52 by at least approximately 70%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:52 by at least approximately 75%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:52 by at least approximately 80%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:52 by at least approximately 85%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 86% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 87% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 88% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 89% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 90% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 91% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 92% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 93% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 94% identical to that of SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 95% identical to that of SEQ ID NO:52.In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:52. In some embodiments, the IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:52.

[0261] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence exhibiting at least approximately 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:53 by at least approximately 70%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:53 by at least approximately 75%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:53 by at least approximately 80%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:53 by at least approximately 85%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 86% identical to that of SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 87% identical to that of SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 88% identical to that of SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 89% identical to that of SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 90% identical to that of SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 91% identical to that of SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 92% identical to that of SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 93% identical to that of SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:53.In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:53. In some embodiments, the IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:53.

[0262] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence exhibiting at least approximately 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:54 by at least approximately 70%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:54 by at least approximately 75%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:54 by at least approximately 80%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:54 by at least approximately 85%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 86% identical to that of SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 87% identical to that of SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 88% identical to that of SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 89% identical to that of SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 90% identical to that of SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 91% identical to that of SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 92% identical to that of SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 93% identical to that of SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:54.In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:54. In some embodiments, the IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:54.

[0263] In some embodiments, the mutation is a deletion, and the variant is a truncated IL-9R intracellular signaling domain shortened by the deletion of one or more amino acid residues at either the N-terminus or C-terminus or both of the wild-type IL-9R intracellular domain. In some embodiments, the truncated IL-9R is a sequence of at least 150 amino acids in length derived from the wild-type IL-9R intracellular signaling domain shown at SEQ ID NO:8, shortened by the deletion of one or more amino acid residues at either the N-terminus or C-terminus or both of the SEQ ID NO:8. In some embodiments, the truncated IL-9R signaling domain is a sequence of 150 to 229 amino acids of the sequence shown at SEQ ID NO:8. In some embodiments, the variant IL-9R is an N-terminal truncation of up to 80 amino acids. In some embodiments, the variant IL-9R is an N-terminal truncation of up to 70 amino acids. In some embodiments, the variant IL-9R is an N-terminal truncation of up to 680 amino acids. In some embodiments, the variant IL-9R is N-terminally shortened by up to 50 amino acids. In some embodiments, the variant IL-9R is N-terminally shortened by up to 40 amino acids. In some embodiments, the variant IL-9R is N-terminally shortened by up to 30 amino acids. In some embodiments, the variant IL-9R is N-terminally shortened by up to 20 amino acids. In some embodiments, the variant IL-9R is N-terminally shortened by up to 10 amino acids. In some embodiments, the variant IL-9R is N-terminally shortened by up to 7 amino acids. In some embodiments, the variant IL-9R is N-terminally shortened by up to 5 amino acids. In some embodiments, one or more further mutations may be present in any of the such shortened IL-9R intracellular domains.

[0264] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence exhibiting at least approximately 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:55 by at least approximately 70%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:55 by at least approximately 75%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:55 by at least approximately 80%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:55 by at least approximately 85%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 86% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 87% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 88% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 89% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:55.In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO: 55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO: 55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO: 55. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO: 55. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO: 55. In some embodiments, the IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 55.

[0265] In some embodiments, the IL-9R intracellular domain contains an amino acid sequence exhibiting at least approximately 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity with SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:56 by at least approximately 70%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:56 by at least approximately 75%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:56 by at least approximately 80%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence identical to SEQ ID NO:56 by at least approximately 85%. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 86% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 87% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 88% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 89% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 90% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 91% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 92% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 93% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 94% identical to SEQ ID NO:56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 95% identical to SEQ ID NO:56.In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 96% identical to SEQ ID NO: 56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 97% identical to SEQ ID NO: 56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 98% identical to SEQ ID NO: 56. In some embodiments, the IL-9R intracellular domain contains an amino acid sequence that is at least approximately 99% identical to SEQ ID NO: 56. In some embodiments, the IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO: 56. In some embodiments, the IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 56. Examples of such intracellular domains, including wild-type or variant IL-9R intracellular domains or chimeric JAK / STAT fusion intracellular domains, are described in the following subsections.

[0266] In some embodiments, the intracellular domains provided herein may include any of the intracellular domains shown in Table 3. In some cases, the N-terminal and / or C-terminal amino acids adjacent to a given intracellular domain may be included in or deleted from the sequence of the synthetic cytokine receptor, as long as the resulting receptor can constitutively multimerize (e.g., dimerize) to result in constitutive activation of downstream signaling molecules. Therefore, it should be understood that the examples of SEQ ID NOs in Table 3 should not be interpreted as limiting. For example, a particular intracellular domain may be several amino acids longer or shorter than the amino acid sequence shown in its respective SEQ ID NO, for example, 1 to 10, for example, 1, 2, 3, 4, 5, 6, or 7 amino acids longer or shorter. Variants of such SEQ ID NO (e.g., sequences exhibiting at least approximately 90% identity, at least approximately 91% identity, at least approximately 92% identity, at least approximately 93% identity, at least approximately 94% identity, at least approximately 95% identity, e.g., at least approximately 96% identity, 97% identity, 98% identity, or 99% identity to any of SEQ ID NO: 8 or 51-56) are also provided, and synthetic cytokine receptors resulting from containing such variant IL-9R intracellular domains can constitutively multimerize (e.g., dimerize) to lead to constitutive activation of downstream signaling molecules.

[0267] (Table 3) Exemplary intracellular domains TIFF2026525229000016.tif17128

[0268] a. Wild-type IL-9R intracellular domain (ICD) In some embodiments, the intracellular domain is capable of interleukin-9 receptor (IL-9R) signaling. In some embodiments, the IL-9R signaling-capable intracellular domain includes the intracellular domain of IL-9R. In some embodiments, the IL-9R intracellular domain includes naturally occurring IL-9R (i.e., wild-type IL-9R). In some embodiments, the IL-9R intracellular domain includes mammalian IL-9R. In some embodiments, the IL-9R intracellular domain includes human IL-9R. In some embodiments, the IL-9R intracellular domain includes wild-type human IL-9R.

[0269] In some embodiments, the IL-9R intracellular domain is 230 amino acids long. In some embodiments, the wild-type IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:8. In some embodiments, the wild-type IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:8. In some embodiments, the wild-type IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:8.

[0270] b. Variant IL-9R intracellular domain (ICD) In some embodiments, the intracellular domain is an interleukin-9 receptor (IL-9R) signaling-capable variant IL-9R intracellular domain. In some embodiments, the variant IL-9R intracellular domain contains one or more mutations compared to naturally occurring IL-9R. The one or more mutations may be substitutions, insertions, deletions, or combinations thereof. In some embodiments, the one or more mutations promote downstream signaling. Methods for determining whether a particular mutation induces downstream signaling are known to those skilled in the art (e.g., assays for phosphorylation of STAT1, STAT3, or STAT5 after proliferation of receptor-possessing cells tested in the absence of growth factors). In some embodiments, the variant IL-9R intracellular domain induces signaling through the STAT1 pathway, the STAT3 pathway, and / or the STAT5 pathway.

[0271] In some embodiments, STAT1 signaling by variant IL-9R is increased compared to STAT1 signaling by wild-type IL-9R. In some embodiments, STAT3 signaling by variant IL-9R is increased compared to STAT3 signaling by wild-type IL-9R. In some embodiments, STAT5 signaling by variant IL-9R is increased compared to STAT5 signaling by wild-type IL-9R. In some embodiments, signaling through the STAT1, STAT3, and / or STAT5 pathways persists for a longer period compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R. In some embodiments, the duration is approximately 6 hours, 12 hours, 24 hours, 36 hours, 48 ​​hours, 72 hours, or longer. In some embodiments, the duration is approximately 24 hours. In some embodiments, signaling through STAT1, STAT3, and / or STAT5 persists as long as the cell expresses one of the synthetic cytokine receptors provided herein. In some embodiments, sustained STAT1, STAT3, and / or STAT5 signaling is determined by the phosphorylation state of STAT1, STAT3, and / or STAT5.

[0272] In some embodiments, the IL-9R intracellular domain includes deletions of regions such as those outlined in Table 4. In some embodiments, the IL-9R intracellular domain includes deletions of consecutive amino acids compared to the wild-type IL-9R intracellular domain. In some embodiments, the IL-9R intracellular domain includes deletions of regions such as those outlined in Table 4 at the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the IL-9R intracellular domain includes deletions of regions at the N-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the IL-9R intracellular domain includes deletions of consecutive amino acid regions within the wild-type IL-9R intracellular domain. In some embodiments, the IL-9R intracellular domain includes deletions of discontinuous amino acids compared to the wild-type IL-9R intracellular domain. In some embodiments, the IL-9R intracellular domain includes deletions of multiple regions of the wild-type IL-9R intracellular domain, such as those outlined in Table 4. Exemplary deletion regions from the IL-9R intracellular domain are shown in Table 4A.

[0273] (Table 4A) Exemplary deletion mutations TIFF2026525229000017.tif107142

[0274] Exemplary deletion regions from the IL-9R intracellular domain are shown in Table 4B. In some embodiments, the IL-9R intracellular domain is missing a sequence of amino acids located at 29-38, 39-48, 69-78, or 154-163 relative to the amino acid numbering of SEQ ID NO:8, or a sequence of amino acids containing these, or any combination thereof.

[0275] (Table 4B) Exemplary deletion areas TIFF2026525229000018.tif45128

[0276] In some embodiments, the IL-9R intracellular domain contains deletions of multiple regions from the wild-type IL-9R intracellular domain. Exemplary intracellular domains containing deletions of multiple regions from the IL-9R intracellular domain are shown in Table 4C.

[0277] (Table 4C) Exemplary deletion of multiple regions TIFF2026525229000019.tif119142

[0278] In some embodiments, the variant IL-9R intracellular domain is a shortened form of IL-9R lacking a continuous amino acid sequence between the N-terminus and C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the variant IL-9R intracellular domain is a shortened form of IL-9R lacking a discontinuous amino acid sequence between the N-terminus and C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the variant IL-9R intracellular domain is a shortened form of IL-9R lacking a continuous amino acid sequence at the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the variant IL-9R intracellular domain is a shortened form of IL-9R lacking a continuous amino acid sequence at the N-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the variant IL-9R intracellular domain is a shortened form of IL-9R lacking a discontinuous amino acid sequence at the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the variant IL-9R intracellular domain is a shortened form of IL-9R lacking a discontinuous amino acid sequence at the N-terminus of the wild-type IL-9R intracellular domain. In some of these embodiments, the shortening is located at the C-terminus of the wild-type IL-9R intracellular domain, indicated by SEQ ID NO:8. In some of these embodiments, the shortening is located at the N-terminus of the wild-type IL-9R intracellular domain, indicated by SEQ ID NO:8.

[0279] In some embodiments, the truncated IL-9R intracellular domain or its variant is shortened by 62 to 99 consecutive amino acids from the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the truncated IL-9R intracellular domain or its variant is shortened by 61 consecutive amino acids from the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the truncated IL-9R intracellular domain or its variant is shortened by 62 consecutive amino acids from the C-terminus of the wild-type IL-9R intracellular domain. In some embodiments, the truncated IL-9R intracellular domain or its variant is shortened by 99 consecutive amino acids from the C-terminus of the wild-type IL-9R intracellular domain.

[0280] In some embodiments, the variant IL-9R intracellular domain is a shortened form of IL-9R lacking amino acids 132-230 of SEQ ID NO:8. In some embodiments, the variant IL-9R intracellular domain lacks amino acids 134-230 of SEQ ID NO:8.

[0281] In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:84. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:84. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:84. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:85. In some embodiments, the variant IL-9R intracellular domain includes the amino acid sequence shown in SEQ ID NO:85. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:85. In some embodiments, the variant IL-9R intracellular domain includes an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:51. In some embodiments, the variant IL-9R intracellular domain includes the amino acid sequence shown in SEQ ID NO:51. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:51. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:52. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:52.In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 52. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO: 86. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO: 86. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 86. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:87. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:87. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:87. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:53. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:53. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 53. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO: 88. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO: 88. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 88. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:89. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:89. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:89. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:90. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:90. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:90. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:91.In some embodiments, the variant IL-9R intracellular domain includes the amino acid sequence shown in SEQ ID NO:91. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:91. In some embodiments, the variant IL-9R intracellular domain includes an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:92. In some embodiments, the variant IL-9R intracellular domain includes the amino acid sequence shown in SEQ ID NO:92. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:92. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:93. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:93. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:93. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:94. In some embodiments, the variant IL-9R intracellular domain comprises the amino acid sequence shown in SEQ ID NO:94. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:94.In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:54. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:54. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:54. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:95. In some embodiments, the variant IL-9R intracellular domain includes the amino acid sequence shown in SEQ ID NO:95. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:95. In some embodiments, the variant IL-9R intracellular domain includes an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:96. In some embodiments, the variant IL-9R intracellular domain includes the amino acid sequence shown in SEQ ID NO:96. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:96. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:97. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:97.In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:97. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:98. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:98. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:98. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:99. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:99. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:99. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:100. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:100. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:100. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:55.In some embodiments, the variant IL-9R intracellular domain includes the amino acid sequence shown in SEQ ID NO: 55. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 55. In some embodiments, the variant IL-9R intracellular domain includes an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO: 56. In some embodiments, the variant IL-9R intracellular domain includes the amino acid sequence shown in SEQ ID NO: 56. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO: 56. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:103. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:103. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:103. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:104. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:104. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:104.

[0282] In some embodiments, the variant IL-9R intracellular domain includes one or more amino acid deletions and insertions. In some embodiments, the IL-9R intracellular domain includes deletions and insertions of consecutive amino acid sequences. In some embodiments, the length of the deleted amino acid sequence and the length of the inserted amino acid sequence are the same. In some embodiments, the lengths of the deleted amino acid sequence and the inserted amino acid sequence are different.

[0283] In some embodiments, the variant IL-9R intracellular domain includes substitution of one or more STAT-binding domains in a sequence of amino acids. In some embodiments, the sequence of amino acids is substituted with a single STAT-binding domain. In some embodiments, the STAT-binding domain is derived from a type I cytokine receptor. In some embodiments, the type I cytokine receptor is selected from the group consisting of interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R), interleukin-7 receptor (IL-7R), interleukin-13 receptor (IL-13R), interleukin-15 receptor (IL-15R), and interleukin-2 receptor (IL-21R). In some embodiments, the type I cytokine receptor is IL-2R.

[0284] In some embodiments, a sequence of amino acids is substituted with one or more STAT-binding domains. In some embodiments, a sequence of amino acids is substituted with one STAT-binding domain. In some embodiments, a sequence of amino acids is substituted with two STAT-binding domains. In some embodiments, a sequence of amino acids is substituted with three STAT-binding domains. In some embodiments, a sequence of amino acids is substituted with four STAT-binding domains. In some embodiments, a sequence of amino acids is substituted with five STAT-binding domains. In some embodiments, substitution of a sequence of amino acids with one, two, three, four, or five STAT-binding domains increases STAT binding and signal transduction.

[0285] In some embodiments, the variant IL-9R intracellular domain includes one or more YLPQ, YRPQ, YLPL, or YLKQ STAT-binding domains. In some embodiments, the variant IL-9R intracellular domain includes two or more YLPQ, YRPQ, YLPL, or YLKQ STAT-binding domains. In some embodiments, the variant IL-9R intracellular domain includes three or more YLPQ, YRPQ, YLPL, or YLKQ STAT-binding domains. In some embodiments, the variant IL-9R intracellular domain includes four or more YLPQ, YRPQ, YLPL, or YLKQ STAT-binding domains. In some embodiments, the variant IL-9R intracellular domain includes five or more YLPQ, YRPQ, YLPL, or YLKQ STAT-binding domains.

[0286] In some embodiments, one or more STAT binding domains include YLPQ (SEQ ID NO: 171). In some embodiments, one or more STAT binding domains include YRPQ (SEQ ID NO: 172). In some embodiments, one or more STAT binding domains include YLPL (SEQ ID NO: 173). In some embodiments, one or more STAT binding domains include YLKQ (SEQ ID NO: 174). In some embodiments, the STAT binding domain includes a STAT1 binding domain. In some embodiments, the STAT binding domain includes a STAT3 binding domain. In some embodiments, the STAT binding domain includes a STAT5 binding domain.

[0287] In some embodiments, the STAT5 binding domain is 11 amino acids long. In some embodiments, the STAT5 binding domain contains the amino acid sequence LNTDAYLSLQE (SEQ ID NO: 120). In some embodiments, the sequence of amino acids substituted for one or more STAT binding domains contains 11 amino acid residues. In some embodiments, the sequence of amino acids substituted for one or more STAT binding domains contains SNNNNYCALGC (SEQ ID NO: 170). Exemplary insertion mutations with one STAT binding domain, two STAT binding domains, and three STAT binding domains are shown in Table 5.

[0288] (Table 5) Exemplary insertion mutations TIFF2026525229000020.tif32142

[0289] In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:110. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:110. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:110. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:111. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:111. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:111. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:112. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:112. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:112.

[0290] In some embodiments, the variant IL-9R intracellular domain contains one or more amino acid substitutions relative to the wild-type IL-9R intracellular domain (SEQ ID NO: 8). In some embodiments, the variant IL-9R intracellular domain contains a STAT binding motif. In some embodiments, one or more amino acid substitutions are located at the STAT binding motif of the IL-9R intracellular domain. In some embodiments, the STAT binding motif contains a STAT1 binding motif, a STAT3 binding motif, and / or a STAT5 binding motif.

[0291] In some embodiments, the STAT binding motif includes YLPQ (SEQ ID NO: 171). In some embodiments, the STAT binding motif includes a variant STAT binding motif. In some embodiments, the variant STAT binding motif includes one or more amino acid substitutions relative to SEQ ID NO: 171. In some embodiments, the amino acid substitution includes the substitution of an arginine residue for a leucine residue relative to SEQ ID NO: 171. In some embodiments, the amino acid substitution includes the substitution of a leucine residue for a glutamine residue relative to SEQ ID NO: 171. In some embodiments, the amino acid substitution includes the substitution of proline for lysine relative to SEQ ID NO: 171.

[0292] In some embodiments, the STAT binding motif includes YRPQ (SEQ ID NO: 172). In some embodiments, the STAT binding motif includes YLPL (SEQ ID NO: 173). In some embodiments, the STAT binding motif includes YLKQ (SEQ ID NO: 174).

[0293] In some embodiments, STAT1, STAT3, and / or STAT5 are linked to YLPQ. In some embodiments, STAT1, STAT3, and STAT5 are linked to YLPQ. In some embodiments, STAT1, and / or STAT3 are linked to YRPQ. In some embodiments, STAT1 and STAT3 are linked to YRPQ. In some embodiments, STAT5 is linked to YLPL. In some embodiments, STAT1, STAT3, and / or STAT5 are linked to YLKQ. In some embodiments, STAT1, STAT3, and STAT5 are linked to YLKQ.

[0294] In some embodiments, the substitution of one or more STAT binding motifs relative to SEQ ID NO:171(YLPQ) is shown in Table 6.

[0295] (Table 6) Exemplary substitution mutations TIFF2026525229000021.tif38142

[0296] In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:107. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:107. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:107. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least approximately 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO:108. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:108. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:108. In some embodiments, the variant IL-9R intracellular domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:109. In some embodiments, the variant IL-9R intracellular domain contains the amino acid sequence shown in SEQ ID NO:109. In some embodiments, the variant IL-9R intracellular domain consists of the amino acid sequence shown in SEQ ID NO:109. In some embodiments, the variant IL-9R intracellular domain provided herein may contain any of the variant IL-9R intracellular domains shown in Table 7. In some embodiments, the variant IL-9R intracellular domain may contain any combination of the variant IL-9R intracellular domains shown in Table 7.In some embodiments, the variant IL-9R intracellular domain comprises one or more amino acid deletions and one or more amino acid substitutions relative to the wild-type IL-9R intracellular domain. In some embodiments, the variant IL-9R intracellular domain comprises one or more amino acid deletions and one or more amino acid insertions relative to the wild-type IL-9R intracellular domain.

[0297] In specific embodiments, the variant IL-9R intracellular domain includes one or amino acid substitutions represented by IL9R(large D2)(SEQ ID NO:103), IL9R(large D2')(SEQ ID NO:104), or IL9R(d15)(SEQ ID NO:54), and IL9R(Mut6YRPQ)(SEQ ID NO:107), IL9R(Mut7YLPL)(SEQ ID NO:108), or IL9R(Mut9YLKQ)(SEQ ID NO:109).

[0298] (Table 7) Exemplary variant IL-9R intracellular domain TIFF2026525229000022.tif13128

[0299] 2. Chimeric JAK / STAT fusion intracellular domain (ICD) In some embodiments, the intracellular domain is a chimeric fusion domain capable of interleukin-9 receptor (IL-9R) signaling. In some embodiments, the chimeric fusion domain includes a JAK-binding domain derived from IL-7R fused with a STAT-binding domain derived from IL-9R. In some embodiments, the chimeric fusion domain includes a JAK-binding domain derived from TpoR fused with a STAT-binding domain derived from IL-9R.

[0300] In some embodiments, chimeric JAK / STAT fusion domains promote downstream signaling. Methods for determining whether a chimeric JAK / STAT fusion domain induces downstream signaling are known to those skilled in the art (e.g., assays for phosphorylation of STAT1, STAT3, or STAT5 after proliferation of receptor-possessing cells tested in the absence of growth factors). In some embodiments, chimeric JAK / STAT fusion domains induce signaling through the STAT1 pathway, the STAT3 pathway, and / or the STAT5 pathway.

[0301] In some embodiments, STAT1 signaling via the chimeric JAK / STAT fusion domain is increased compared to STAT1 signaling via wild-type IL-9R. In some embodiments, STAT3 signaling via the chimeric JAK / STAT fusion domain is increased compared to STAT3 signaling via wild-type IL-9R. In some embodiments, STAT5 signaling via the chimeric JAK / STAT fusion domain is increased compared to STAT5 signaling via wild-type IL-9R. In some embodiments, STAT1, STAT3, and / or STAT5 signaling via the chimeric JAK / STAT fusion domain persists for a longer period compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R. In some embodiments, the duration is approximately 6 hours, 12 hours, 24 hours, 36 hours, 48 ​​hours, or 72 hours. In some embodiments, the duration is approximately 24 hours. In some embodiments, the sustained STAT1, STAT3, and / or STAT5 signaling is determined by the phosphorylation state of STAT1, STAT3, and / or STAT5.

[0302] In some embodiments, the chimeric JAK / STAT fusion domain comprises a JAK-binding domain derived from a type I cytokine receptor and a STAT-binding domain derived from an IL-9R intracellular domain.

[0303] In some embodiments, the IL-9R STAT-binding domain contains amino acid residues 73-230 of SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain is 59-158 amino acids long and contains an IL-9R STAT-binding motif. In some embodiments, the IL-9R STAT-binding motif contains YLPQ (SEQ ID NO:171). In some embodiments, the IL-9R STAT-binding domain contains one or more amino acid deletions relative to the IL-9R ICD shown in SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking a continuous amino acid sequence at the N-terminus of SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking a continuous amino acid sequence at the C-terminus of SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain is a truncated IL-9R STAT-binding domain lacking amino acids at positions 1-72 of SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain is a truncated IL-9R STAT-binding domain lacking amino acids at positions 132-230 of SEQ ID NO:8. In some embodiments, the IL-9R STAT-binding domain is a truncated IL-9R STAT-binding domain lacking amino acids at positions 1-72 and 132-230 of SEQ ID NO:8. Exemplary STAT-binding domains derived from the IL-9R receptor are shown in Table 8.

[0304] (Table 8) Exemplary JAK binding domains TIFF2026525229000023.tif30152

[0305] In some embodiments, the IL-9R STAT-binding domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:122. In some embodiments, the IL-9R STAT-binding domain contains the amino acid sequence of SEQ ID NO:122. In some embodiments, the IL-9R STAT-binding domain consists of the amino acid sequence shown in SEQ ID NO:122. In some embodiments, the IL-9R STAT-binding domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:123. In some embodiments, the IL-9R STAT-binding domain contains the amino acid sequence of SEQ ID NO:123. In some embodiments, the IL-9R STAT-binding domain consists of the amino acid sequence shown in SEQ ID NO:123.

[0306] In some embodiments, the type I cytokine receptor is selected from the group consisting of interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R), interleukin-7 receptor (IL-7R), interleukin-13 receptor (IL-13R), interleukin-15 receptor (IL-15R), interleukin-2 receptor (IL-21R), and thrombopoietin receptor (TpoR). In some embodiments, the type I cytokine receptor is IL-7R. In some embodiments, the type I cytokine receptor is TpoR. Exemplary JAK-binding domains derived from type I cytokine receptors are shown in Table 9.

[0307] (Table 9) Exemplary JAK binding domains TIFF2026525229000024.tif51152

[0308] In some embodiments, the IL-7R JAK-binding domain is up to 65 amino acids long. In some embodiments, the IL-7R JAK-binding domain is 65 amino acids long. In some embodiments, the IL-7R JAK-binding domain contains a box 1 motif. In some embodiments, the IL-7R JAK-binding domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:119. In some embodiments, the IL-7R JAK-binding domain contains the amino acid sequence of SEQ ID NO:119. In some embodiments, the IL-7R JAK-binding domain consists of the amino acid sequence shown in SEQ ID NO:119. In some embodiments, the IL-7R JAK-binding domain contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:121. In some embodiments, the IL-7R JAK-binding domain contains the amino acid sequence of SEQ ID NO:121. In some embodiments, the IL-7R JAK-binding domain consists of the amino acid sequence shown in SEQ ID NO:121.

[0309] In some embodiments, the chimeric JAK / STAT fusion intracellular domain comprises a fusion of either the JAK-binding domain or the STAT-binding domain. In some embodiments, the chimeric JAK / STAT fusion intracellular domain contains approximately 223 amino acids. In some embodiments, the JAK / STAT fusion intracellular domain is a shortened version of the 223-amino acid sequence. In some embodiments, the JAK / STAT fusion intracellular domain is shortened at the C-terminus. In some embodiments, 99 amino acids are cleaved from the C-terminus. In other embodiments, 97 amino acids are cleaved from the C-terminus. In some embodiments, the chimeric JAK / STAT fusion intracellular domain provided herein may comprise any of the intracellular domains shown in Table 10.

[0310] (Table 10) Exemplary chimeric JAK / STAT fusion intracellular domains TIFF2026525229000025.tif13128

[0311] In some embodiments, the chimeric JAK / STAT fusion contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:114. In some embodiments, the chimeric JAK / STAT fusion contains the amino acid sequence shown in SEQ ID NO:114. In some embodiments, the chimeric JAK / STAT fusion consists of the amino acid sequence shown in SEQ ID NO:114. In some embodiments, the chimeric JAK / STAT fusion contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:116. In some embodiments, the chimeric JAK / STAT fusion comprises the amino acid sequence shown in SEQ ID NO:116. In some embodiments, the chimeric JAK / STAT fusion consists of the amino acid sequence shown in SEQ ID NO:116. In some embodiments, the TpoR JAK binding domain comprises a variant TpoR JAK binding domain. In some embodiments, the variant TpoR JAK binding domain The TpoR JAK-binding domain contains one or more mutations compared to the wild-type TpoR JAK-binding domain shown in TIFF2026525229000026.tif16146. In some embodiments, one or more mutations promote homodimerization of the synthetic cytokine receptor. In some embodiments, one or more mutations promote α-helix dimerization. In some embodiments, one or more mutations substitute at least one amino acid residue in the wild-type TpoR JAK-binding domain with an arginine residue. In some embodiments, the TpoR JAK-binding domain is up to 70 amino acids long. In some embodiments, the TpoR JAK-binding domain is 69 amino acids long. In some embodiments, the TpoR JAK-binding domain contains a box-1 motif. In some embodiments, the TpoR JAK-binding domain contains an amino acid sequence that is at least approximately 85% identical to SEQ ID NO:175. In some embodiments, the TpoR JAK-binding domain contains the amino acid sequence of SEQ ID NO:175. In some embodiments, the TpoR JAK-binding domain consists of the amino acid sequence shown in SEQ ID NO:175. In some embodiments, the box 1 motif contains the amino acid sequence LWPSLPDLH (SEQ ID NO:176).

[0312] D. Variants of synthetic cytokine receptors Amino acid sequence modifications of synthetic cytokine receptors provided herein are also intended. For example, it may be desirable to improve signal transduction induced by synthetic cytokine receptors; it may also be desirable to improve other biological properties of synthetic cytokine receptors, including, but not limited to, thermal stability, expression levels, or solubility. Therefore, it is intended that variants may also be prepared in addition to the synthetic cytokine receptors described herein.

[0313] In some embodiments, the synthetic cytokine receptors provided herein are chemically modified, for example, by covalent attachment of any type of molecule to the synthetic cytokine receptor. Exemplary, non-limiting modifications include glycosylation, acetylation, PEGylation, phosphorylation, amidation, derivatization with known protective / blocking groups, proteolysis, and linkage with cellular ligands or other proteins. Furthermore, the synthetic cytokine receptor may contain one or more non-classical amino acids.

[0314] In some embodiments, the variation may be a substitution, deletion, or insertion of one or more codons encoding the synthetic cytokine receptor, resulting in a change in the amino acid sequence compared to the initial sequence.

[0315] Amino acid substitutions may be the result of substitutions between amino acids having similar structural and / or chemical properties, such as the substitution of leucine for serine, or other conservative amino acid substitutions. Standard techniques known to those skilled in the art, including site-directed mutagenesis and PCR-mediated mutagenesis resulting in amino acid substitutions, may be used to introduce mutations into the nucleotide sequences encoding the molecules provided herein.

[0316] A "conservative amino acid substitution" is a substitution in which amino acid residues having similarly charged side chains are substituted for each other. Families of amino acid residues having similarly charged side chains have been clearly defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with non-charged side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), amino acids with nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), amino acids with β-branched side chains (e.g., threonine, valine, isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Naturally occurring residues can be classified based on common side-chain properties: (1) hydrophobic: norleucine, Met, Ala, Val, Leu, lie; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gin; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues that affect chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe. Alternatively, for example, mutations may be randomly introduced into all or part of the coding sequence by saturation mutagenesis, and the resulting mutants may be screened for biological activity to identify mutants that retain activity. After mutagenesis, the encoded protein can be expressed and its activity can be determined.

[0317] Substantial modification of the biological properties of synthetic cytokine receptors is achieved by selecting substitutions that have significantly different effects on maintaining (a) the structure of the polypeptide scaffold in the substitution region, e.g., the three-dimensional structure of sheets or helices, (b) the molecular charge or hydrophobicity at the target site, or (c) the bulk of the side chains.

[0318] Amino acid sequence insertions include fusions at the amino and / or carboxyl termini, ranging in length from one residue to polypeptides containing 100 or more residues, as well as intrasequence insertions of single or multiple amino acid residues.

[0319] Amino acid sequence deletions include deletions at the amino and / or carboxyl terminals of polypeptides ranging in length from one residue to 100 or more residues, as well as intrasequence deletions of single or multiple amino acid residues.

[0320] E. Exemplary synthetic cytokine receptor In some embodiments, the synthetic cytokine receptors provided herein may comprise any of the extracellular domain, transmembrane domain, and intracellular domain provided herein. In some embodiments, the synthetic cytokine receptors provided herein may comprise one extracellular domain from Table 1, one transmembrane domain from Table 2, and one intracellular domain from any of Tables 3-5, 7, and 10. In some embodiments, the synthetic cytokine receptor may comprise any combination of one extracellular domain, one transmembrane domain, and one intracellular domain. Various combinations of extracellular domains, transmembrane domains, and intracellular domains provided in exemplary synthetic cytokine receptors are shown in Table 11.

[0321] In some embodiments, the synthetic cytokine receptor may include an extracellular domain derived from the CD34 receptor. In some embodiments, the synthetic cytokine receptor may include an extracellular domain derived from the DAP12 receptor. In some embodiments, the synthetic cytokine receptor may include an extracellular domain derived from the GpA receptor. In some embodiments, the extracellular domain derived from GpA includes the amino acid sequence of SEQ ID NO: 16. In some embodiments, the synthetic cytokine receptor may include an extracellular domain derived from a further shortened GpA receptor (i.e., tGpA in Table 11). In some embodiments, the shortened GpA extracellular domain includes the amino acid sequence of SEQ ID NO: 17. In some embodiments, the synthetic cytokine receptor may include an extracellular domain derived from the CD8 receptor. In some embodiments, the extracellular domain derived from CD8 includes the amino acid sequence of SEQ ID NO: 185. In some embodiments, the synthetic cytokine receptor may include an extracellular domain derived from a further shortened CD8 receptor (i.e., tCD8 in Table 11). In some embodiments, the shortened CD8 extracellular domain includes the amino acid sequence of SEQ ID NO: 19. In some embodiments, the synthetic cytokine receptor may include an extracellular domain derived from the Muc24 receptor.

[0322] In some embodiments, the synthetic cytokine receptor may include a transmembrane domain derived from the IL7R receptor. In some embodiments, the transmembrane domain derived from IL7R may be a variant of wild-type IL7R based on SEQ ID NO:71 (e.g., IL7R in Table 11). * and IL7R *2) Includes. In some embodiments, the variant of wild-type IL-7R (SEQ ID NO: 71) includes one or more amino acid insertions. In some embodiments, the variant of wild-type IL-7R includes a trimer peptide. In some embodiments, the trimer includes cysteine. In some embodiments, the trimer includes proline. In some embodiments, the trimer includes threonine. In some embodiments, the trimer peptide includes CPT. In some embodiments, the variant of IL7R includes the amino acid sequence shown in SEQ ID NO: 6 (i.e., IL7R in Table 11) * ) includes. In some embodiments, variants of IL7R further include one or more amino acid substitutions and insertions in the C-terminal domain based on SEQ ID NO:6 or SEQ ID NO:71. In some embodiments, one or more amino acid substitutions include a W to K substitution. In some embodiments, one or more amino acid insertions include a K insertion. In some embodiments, one or more amino acid insertions include an R insertion. In some embodiments, one or more amino acid insertions include an I insertion. In some embodiments, one or more amino acid insertions include K, R, and I insertions. In some embodiments, variants of IL7R include the amino acid sequence shown in SEQ ID NO:7 (i.e., IL7R in Table 11) * Includes 2).

[0323] In some embodiments, the synthetic cytokine receptor may include a transmembrane domain derived from the Muc24 receptor.

[0324] In some embodiments, synthetic cytokine receptors may include transmembrane domains derived from TACI receptors.

[0325] In some embodiments, the synthetic cytokine receptor may include a transmembrane domain derived from the CPT1 receptor.

[0326] In some embodiments, synthetic cytokine receptors may include a transmembrane domain derived from the DR5 receptor.

[0327] In some embodiments, the synthetic cytokine receptor may include a transmembrane domain derived from the GpA receptor.

[0328] In some embodiments, the synthetic cytokine receptor has a transmembrane domain derived from a modified GpA receptor (tGpA in Table 11). * ) may include. In some embodiments, the modified GpA transmembrane domain contains one or more amino acid deletions. In some embodiments, the modified GpA transmembrane domain contains one or more amino acid deletions at the C-terminus. In some embodiments, the GpA transmembrane domain contains the amino acid sequence of SEQ ID NO:22. In some embodiments, the modified GpA transmembrane domain (i.e., tGPA in Table 11) contains the amino acid sequence of SEQ ID NO:43.

[0329] In some embodiments, the synthetic cytokine receptor may include an intracellular domain derived from the wild-type IL9R receptor. In some embodiments, the synthetic cytokine receptor may include an intracellular domain derived from a further modified wild-type IL9R. In some embodiments, the modification includes the shortening or deletion of one or more amino acid residues. In some embodiments, the modification includes the insertion of one or more amino acid residues. As described in Section II.C.1.a., in some embodiments, the synthetic cytokine receptor may include an intracellular domain derived from the wild-type IL9R (i.e., SEQ ID NO: 8 in Table 11). As described in Section II.C.1.b., in some embodiments, the synthetic cytokine receptor may include an intracellular domain derived from a further shortened wild-type IL9R (i.e., SEQ ID NO: 51-56 in Table 11). As described in Section II.C.2., in some embodiments, the synthetic cytokine receptor may include a chimeric JAK / STAT fusion (i.e., SEQ ID NO: 114, 116, and 181 in Table 11). Furthermore, variants of such SEQ ID NOs (e.g., sequences exhibiting at least approximately 90% identity, at least approximately 91% identity, at least approximately 92% identity, at least approximately 93% identity, at least approximately 94% identity, at least approximately 95% identity, e.g., at least approximately 96% identity, 97% identity, 98% identity, or 99% identity to any of SEQ ID NOs: 31-42, 47-50, and 58-64) are also provided, and synthetic cytokine receptors resulting from containing such variant IL-9R intracellular domains can constitutively multimerize (e.g., dimerize) to lead to constitutive activation of downstream signaling molecules.

[0330] In some embodiments, any of the sequences indicated by their respective SEQ ID NOs may also contain an N-terminal signal sequence. In some embodiments, the signal sequence is a heterologous signal sequence that is not naturally contiguous in the wild-type sequence. In some embodiments, the signal sequence is a native signal sequence that is contiguous with the extracellular domain sequence. In some embodiments, the signal sequence is the CD34 signal peptide. The filename is TIFF2026525229000027.tif4128. In some embodiments, the signal sequence is the DAP12 signal peptide. The filename is TIFF2026525229000028.tif4128.

[0331] In some embodiments, any of the sequences indicated by their respective SEQ ID NOs may further include a tag or other sequence to facilitate the detection of the expressed protein. In some embodiments, the sequence may be a fluorescent moiety or a peptide tag sequence. For example, peptide tags may include a Flag tag, Rho1D4 tag, Myc tag, His tag, CL7 tag, HA tag, or V5 tag. In some embodiments, the sequence is a Flag tag (DYKDDDDK; SEQ ID NO: 3).

[0332] (Table 11) Exemplary IL9R synthetic cytokine receptors TIFF2026525229000029.tif123146TIFF2026525229000030.tif214146TIFF2026525229000031.tif210146t=abbreviated type * = wild type variant

[0333] In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:31. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:31. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:31. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:9. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence of SEQ ID NO:9. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:9. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:65. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:65. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:65. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:10. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:10. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:10.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:66. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:66. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:66. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:11. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:11. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:11. In some embodiments, the synthetic cytokine receptor comprises an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:67. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:67. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:67. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:12. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:12. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:12.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:32. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:32. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:32. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:24. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:24. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:24. In some embodiments, the synthetic cytokine receptor comprises an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:33. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:33. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:33. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:25. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:25. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:25.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:34. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:34. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:34. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:26. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:26. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:26. In some embodiments, the synthetic cytokine receptor comprises an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:35. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:35. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:35. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:27. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:27. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:27.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:36. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:36. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:36. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:28. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:28. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:28. In some embodiments, the synthetic cytokine receptor is... The synthetic cytokine receptor contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:38. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:38. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:38. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:29. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:29. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:29. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:37. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:37. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:37. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:30. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:30. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:30.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:39. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:39. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:39. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least approximately 70%, approximately 75%, approximately 80%, approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, approximately 97%, approximately 98%, or approximately 99% identical to SEQ ID NO:40. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:40. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:40. In some embodiments, the synthetic cytokine receptor comprises an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:41. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:41. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:41. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:42. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:42. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:42.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:47. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:47. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:47. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:48. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:48. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:48. In some embodiments, the synthetic cytokine receptor comprises an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:49. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:49. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:49. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:50. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:50. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:50.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO: 58. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO: 58. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO: 58. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO: 59. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO: 59. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO: 59. In some embodiments, the synthetic cytokine receptor comprises an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO: 60. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO: 60. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO: 60. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:61. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:61. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:61.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:62. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:62. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:62. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:63. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:63. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:63. In some embodiments, the synthetic cytokine receptor comprises an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:113. In some embodiments, the synthetic cytokine receptor comprises the amino acid sequence shown in SEQ ID NO:113. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:113. In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:115. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:115. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:115.In some embodiments, the synthetic cytokine receptor contains an amino acid sequence that is at least about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to SEQ ID NO:117. In some embodiments, the synthetic cytokine receptor contains the amino acid sequence shown in SEQ ID NO:117. In some embodiments, the synthetic cytokine receptor consists of the amino acid sequence shown in SEQ ID NO:117.

[0334] In specific embodiments, the synthetic cytokine receptor pro...

Claims

1. A synthetic cytokine receptor that is a homodimer of the same polypeptide chain, each containing an extracellular domain, a transmembrane domain, and an intracellular domain capable of interleukin-9 receptor (IL-9R) signaling.

2. The synthetic cytokine receptor according to claim 1, wherein the IL-9R signaling-capable intracellular domain comprises an IL-9R intracellular domain or a variant thereof.

3. The synthetic cytokine receptor according to claim 1 or claim 2, wherein the intracellular domain capable of IL-9R signaling includes a chimeric JAK / STAT fusion domain.

4. A synthetic cytokine receptor comprising an extracellular domain, a transmembrane domain, and an intracellular domain of the interleukin-9 receptor (IL-9R) or a variant thereof, wherein the synthetic cytokine receptor is a constitutively active cytokine receptor.

5. A synthetic cytokine receptor according to any one of claims 1 to 4, which is a polymer.

6. A synthetic cytokine receptor according to any one of claims 2 to 5, wherein the extracellular domain, the transmembrane domain, and the IL-9R intracellular domain or a variant thereof are each a polymer of the same polypeptide chain.

7. The synthetic cytokine receptor according to claim 5 or 6, wherein the polymer is a dimer.

8. The synthetic cytokine receptor according to claim 7, wherein the dimer is a homodimer.

9. A synthetic cytokine receptor according to any one of claims 1 to 8, wherein each polypeptide chain is constitutively polymerized.

10. A synthetic cytokine receptor according to any one of claims 1 to 9, comprising at least one self-assembling domain.

11. The synthetic cytokine receptor according to claim 10, wherein the at least one self-assembling domain is the extracellular domain and / or the transmembrane domain.

12. The synthetic cytokine receptor according to any one of claims 1 to 11, wherein it is multimerized through the transmembrane domain and / or the extracellular domain.

13. A synthetic cytokine receptor according to any one of claims 1 to 12, wherein the receptor is polymerized through the transmembrane domain and the extracellular domain.

14. A synthetic cytokine receptor that is a homodimer of the same polypeptide chain, each containing an extracellular domain, a transmembrane domain, and an intracellular domain of the interleukin-9 receptor (IL-9R) or a variant thereof.

15. The synthetic cytokine receptor according to any one of claims 1 to 14, wherein the extracellular domain and / or the transmembrane domain are heterogeneous with respect to IL-9R.

16. The synthetic cytokine receptor according to any one of claims 1 to 15, wherein the transmembrane domain and the extracellular domain are derived from the same protein.

17. The synthetic cytokine receptor according to any one of claims 1 to 15, wherein the transmembrane domain and extracellular domain are transmembrane domains and extracellular domains derived from different proteins.

18. The synthetic cytokine receptor according to any one of claims 1 to 17, wherein the transmembrane domain has a length of 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 amino acids.

19. The synthetic cytokine receptor according to any one of claims 1 to 18, wherein the transmembrane domain comprises a transmembrane domain derived from glycophorin A (GpA), carnitine palmitoyltransferase 1 (CPT1), tumor necrosis factor receptor (TNFR), or Muc24.

20. The synthetic cytokine receptor according to any one of claims 1 to 18, wherein the transmembrane domain comprises a transmembrane domain derived from a thrombopoietin receptor.

21. The synthetic cytokine receptor according to any one of claims 1 to 20, wherein the transmembrane domain promotes α-helix dimerization.

22. The synthetic cytokine receptor according to any one of claims 1 to 21, wherein the transmembrane domain comprises motif GXXXG (SEQ ID NO: 68).

23. The aforementioned transmembrane domain is a motif A synthetic cytokine receptor according to any one of claims 1 to 21, comprising:

24. The synthetic cytokine receptor according to any one of claims 1 to 23, wherein the transmembrane domain is a transmembrane domain derived from glycophorin A (GpA), or a variant thereof comprising one or more mutations (e.g., 1, 2, 3, 4, 5, or 6 mutations) compared to a wild-type GpA transmembrane domain, and the variant GpA is sufficient to promote α-helix dimerization.

25. The synthetic cytokine receptor according to any one of claims 1 to 24, wherein the transmembrane domain comprises a transmembrane domain derived from glycophorin A (GpA).

26. The synthetic cytokine receptor according to any one of claims 1 to 25, wherein the transmembrane domain comprises an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:22 or SEQ ID NO:

43.

27. The synthetic cytokine receptor according to any one of claims 1 to 26, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:22 or SEQ ID NO:

43.

28. The synthetic cytokine receptor according to any one of claims 1 to 22, wherein the transmembrane domain comprises a GXXXG (SEQ ID NO:68) motif and a GXXXA (SEQ ID NO:69) motif.

29. The synthetic cytokine receptor according to any one of claims 1 to 22 and 28, wherein the transmembrane domain comprises a transmembrane domain derived from carnitine palmitoyltransferase 1 (CPT1).

30. The synthetic cytokine receptor according to any one of claims 1 to 22, 28, and 29, wherein the transmembrane domain comprises an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

45.

31. The synthetic cytokine receptor according to any one of claims 1 to 22 and 28 to 30, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

45.

32. A synthetic cytokine receptor according to any one of claims 1 to 21, comprising motif AXXXA (SEQ ID NO: 77) or AXXXS (SEQ ID NO: 78).

33. A synthetic cytokine receptor according to any one of claims 1 to 22 or 32, comprising the motif ΦPXΦ (SEQ ID NO: 75) or ΦTXXAΦ (SEQ ID NO: 76).

34. The synthetic cytokine receptor according to any one of claims 1 to 22, 32, or 33, wherein the transmembrane domain is derived from TNFR.

35. The synthetic cytokine receptor according to claim 34, wherein the TNFR is TACI, DR5, p75NTR, Fas, TNFR1, TNFR2, or OX40.

36. The synthetic cytokine receptor according to any one of claims 1 to 22 and 32 to 35, wherein the transmembrane domain comprises a transmembrane domain derived from DR5.

37. The synthetic cytokine receptor according to any one of claims 1 to 22 and 32 to 36, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

46.

38. The synthetic cytokine receptor according to any one of claims 1 to 22 and 32 to 37, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

46.

39. The synthetic cytokine receptor according to any one of claims 1 to 22 and 32 to 36, wherein the transmembrane domain comprises a transmembrane domain derived from TACI.

40. The synthetic cytokine receptor according to any one of claims 1 to 22, 32 to 36, and 39, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

44.

41. The synthetic cytokine receptor according to any one of claims 1 to 23, 32 to 36, 39, and 40, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

44.

42. The synthetic cytokine receptor according to any one of claims 1 to 19, wherein the transmembrane domain comprises 1 to 6 cysteine ​​residues.

43. The synthetic cytokine receptor according to any one of claims 1 to 19 and 42, wherein the transmembrane domain promotes disulfide bond dimerization.

44. The synthetic cytokine receptor according to claim 43, wherein the disulfide bond dimerization forms 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

45. The synthetic cytokine receptor according to any one of claims 1 to 19 and 42 to 44, wherein the transmembrane domain is a variant transmembrane domain containing one or more mutations compared to the wild-type transmembrane domain for promoting homodimerization of the receptor polypeptide.

46. The synthetic cytokine receptor according to claim 45, wherein the one or more mutations promote α-helix dimerization or disulfide bond dimerization.

47. The synthetic cytokine receptor according to claim 45 or 46, wherein the one or more mutations introduce at least one cysteine ​​into the transmembrane domain.

48. The synthetic cytokine receptor according to any one of claims 45 to 47, wherein the one or more mutations introduce proline into the transmembrane domain.

49. The synthetic cytokine receptor according to any one of claims 45 to 47, wherein the one or more mutations introduce threonine into the transmembrane domain.

50. The synthetic cytokine receptor according to any one of claims 45 to 49, wherein the one or more mutations introduce a trimer peptide of cysteine, proline, and another amino acid other than cysteine ​​and proline into the transmembrane domain.

51. The synthetic cytokine receptor according to any one of claims 45 to 50, wherein the one or more mutations introduce a cysteine-proline-threonine trimer peptide (CPT or TCP) into the transmembrane domain.

52. The transmembrane domain is a variant IL-7R transmembrane domain, and the one or more mutations are wild-type transmembrane sequences. A synthetic cytokine receptor according to any one of claims 45 to 51, located within the same area.

53. The synthetic cytokine receptor according to any one of claims 1 to 19 and 42 to 52, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:

21.

54. The synthetic cytokine receptor according to any one of claims 1 to 19 and 42 to 53, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:

21.

55. The synthetic cytokine receptor according to any one of claims 1 to 19, wherein the transmembrane domain comprises a transmembrane domain derived from Muc24.

56. The synthetic cytokine receptor according to any one of claims 1 to 19 and 55, wherein the transmembrane domain comprises an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

23.

57. The synthetic cytokine receptor according to any one of claims 1 to 19, 55, or 56, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

23.

58. The synthetic cytokine receptor according to any one of claims 1 to 57, wherein the extracellular domain is approximately 150 to 260 amino acids long.

59. The synthetic cytokine receptor according to any one of claims 1 to 58, wherein the extracellular domain is a dimerizing domain.

60. The synthetic cytokine receptor according to claim 59, wherein the dimerization domain includes a hinge region.

61. The synthetic cytokine receptor according to any one of claims 1 to 60, wherein the extracellular domain promotes disulfide bond dimerization.

62. The synthetic cytokine receptor according to any one of claims 1 to 61, wherein the extracellular domain comprises 1 to 6 cysteine ​​residues.

63. The synthetic cytokine receptor according to claim 61 or claim 62, wherein the disulfide bond dimerization forms 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

64. The synthetic cytokine receptor according to any one of claims 1 to 63, wherein the extracellular domain is derived from the extracellular domain of CD34, DAP12, glycophorin A, CD8, or Muc24.

65. The synthetic cytokine receptor according to any one of claims 1 to 63, wherein the extracellular domain comprises an extracellular domain derived from a thrombopoietin receptor.

66. The synthetic cytokine receptor according to any one of claims 1 to 65, wherein the extracellular domain comprises the extracellular domain of CD8 or a truncated portion thereof containing at least one cysteine ​​residue.

67. The synthetic cytokine receptor according to any one of claims 1 to 66, wherein the extracellular domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:18, SEQ ID NO:185, or SEQ ID NO:

19.

68. The synthetic cytokine receptor according to any one of claims 1 to 67, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:18, SEQ ID NO:185, or SEQ ID NO:

19.

69. The synthetic cytokine receptor according to any one of claims 1 to 65, wherein the extracellular domain comprises the extracellular domain of CD34, or a truncated portion thereof containing at least one cysteine ​​residue.

70. The synthetic cytokine receptor according to any one of claims 1 to 65 and 69, wherein the extracellular domain comprises an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

4.

71. The synthetic cytokine receptor according to any one of claims 1 to 65, 69, and 70, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:

4.

72. The synthetic cytokine receptor according to any one of claims 1 to 65, wherein the extracellular domain is the extracellular domain of Muc24, or a truncated portion thereof containing at least one cysteine ​​residue.

73. The synthetic cytokine receptor according to any one of claims 1 to 65 and 72, wherein the extracellular domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

20.

74. The synthetic cytokine receptor according to any one of claims 1 to 65, 72, and 73, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:

20.

75. The synthetic cytokine receptor according to any one of claims 1 to 65, wherein the extracellular domain is the extracellular domain of DAP12, or a truncated portion thereof containing at least one cysteine ​​residue.

76. The synthetic cytokine receptor according to any one of claims 1 to 65 and 75, wherein the extracellular domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

5.

77. The synthetic cytokine receptor according to any one of claims 1 to 65, 75, and 76, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:

5.

78. The synthetic cytokine receptor according to any one of claims 1 to 65, wherein the extracellular domain is the extracellular domain of glycophorin A (GpA), or a truncated portion thereof containing at least one cysteine ​​residue.

79. The synthetic cytokine receptor according to any one of claims 1 to 65 and 78, wherein the extracellular domain comprises an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:16 or SEQ ID NO:

17.

80. The synthetic cytokine receptor according to any one of claims 1 to 65, 78, and 79, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:16 or SEQ ID NO:

17.

81. The synthetic cytokine receptor according to any one of claims 2 to 80, wherein the IL-9R intracellular domain or a variant thereof is approximately 100 to 260 amino acids long.

82. The synthetic cytokine receptor according to any one of claims 2 to 81, wherein the IL-9R intracellular domain or a variant thereof comprises a box 1 motif and / or a box 2 motif.

83. The synthetic cytokine receptor according to any one of claims 2 to 82, wherein the IL-9R intracellular domain or a variant thereof comprises a box 2 motif.

84. The synthetic cytokine receptor according to any one of claims 2 and 4 to 80, wherein the IL-9R intracellular domain or a variant thereof is 230 amino acid long.

85. The synthetic cytokine receptor according to any one of claims 1 to 84, wherein the IL-9R intracellular domain or its variant is the wild-type IL-9R intracellular domain, or a variant thereof containing one or more mutations compared to the wild-type IL-9R intracellular domain shown in SEQ ID NO:

8.

86. The synthetic cytokine receptor according to claim 85, wherein the one or more mutations include the insertion, deletion, and / or substitution of one or more amino acids.

87. The synthetic cytokine receptor according to claim 85 or 86, wherein the one or more mutations promote signaling through the STAT1 pathway, the STAT3 pathway, and / or the STAT5 pathway.

88. The synthetic cytokine receptor according to any one of claims 1 to 87, wherein the IL-9R intracellular domain or a variant thereof contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

8.

89. The synthetic cytokine receptor according to any one of claims 1 to 88, wherein the IL-9R intracellular domain or a variant thereof comprises the amino acid sequence of SEQ ID NO:

8.

90. The synthetic cytokine receptor according to any one of claims 1 to 87, wherein the IL-9R intracellular domain or a variant thereof contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, or SEQ ID NO:

56.

91. The synthetic cytokine receptor according to any one of claims 1 to 87 and 90, wherein the IL-9R intracellular domain or a variant thereof comprises the amino acid sequence of SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, or SEQ ID NO:

56.

92. A synthetic cytokine receptor according to any one of claims 1 to 91, comprising an amino acid sequence that is at least about 85% identical to SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:

67.

93. A synthetic cytokine receptor according to any one of claims 1 to 92, comprising the amino acid sequence of SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:

67.

94. The synthetic cytokine receptor according to any one of claims 2 to 87, wherein the IL-9R intracellular domain or its variant comprises one or more amino acid deletions relative to the wild-type IL-9R intracellular domain (SEQ ID NO: 8).

95. The synthetic cytokine receptor according to any one of claims 2 to 87, wherein the IL-9R intracellular domain or its variant is a truncated IL-9R lacking a continuous amino acid sequence at the C-terminus of the wild-type IL-9R intracellular domain.

96. The synthetic cytokine receptor according to claim 95, wherein the shortened IL-9R intracellular domain or its variant is shortened by 62 to 99 consecutive amino acids from the C-terminus of the wild-type IL-9R intracellular domain.

97. The synthetic cytokine receptor according to any one of claims 2 to 87, wherein the IL-9R intracellular domain or a variant thereof is a shortened IL-9R lacking amino acids 132 to 230 of SEQ ID NO:8 or lacking amino acids 134 to 230 of SEQ ID NO:

8.

98. The synthetic cytokine receptor according to any one of claims 2 to 97, wherein the IL-9R intracellular domain or a variant thereof comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:54, SEQ ID NO:103, or SEQ ID NO:

104.

99. The synthetic cytokine receptor according to any one of claims 2 to 98, wherein the IL-9R intracellular domain or a variant thereof comprises the amino acid sequence of SEQ ID NO: 54, SEQ ID NO: 103, or SEQ ID NO:

104.

100. A synthetic cytokine receptor according to any one of claims 2 to 99, comprising an amino acid sequence that is at least about 85% identical to SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, or SEQ ID NO:

63.

101. A synthetic cytokine receptor according to any one of claims 2 to 100, comprising the amino acid sequence of SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, or SEQ ID NO:

63.

102. The synthetic cytokine receptor according to any one of claims 2 to 87, wherein the IL-9R intracellular domain or its variant comprises one or more amino acid substitutions relative to the wild-type IL-9R intracellular domain (SEQ ID NO: 8).

103. The synthetic cytokine receptor according to claim 102, wherein the IL-9R intracellular domain or a variant thereof comprises a STAT-binding motif or a variant thereof.

104. The synthetic cytokine receptor according to claim 103, wherein the STAT binding motif comprises a STAT1 binding motif, a STAT3 binding motif, and / or a STAT5 binding motif.

105. The synthetic cytokine receptor according to claim 103 or claim 104, wherein the STAT-binding motif comprises YLPQ (SEQ ID NO: 171).

106. The synthetic cytokine receptor according to any one of claims 103 to 105, wherein the STAT binding motif includes a variant STAT binding motif.

107. The synthetic cytokine receptor according to any one of claims 103 to 106, wherein the variant STAT binding motif includes YRPQ (SEQ ID NO: 172).

108. The synthetic cytokine receptor according to any one of claims 103 to 106, wherein the variant STAT binding motif includes YLPL (SEQ ID NO: 173).

109. The synthetic cytokine receptor according to any one of claims 103 to 106, wherein the variant STAT binding motif comprises YLKQ (SEQ ID NO: 174).

110. The synthetic cytokine receptor according to any one of claims 2 to 109, wherein the variant IL-9R intracellular domain or the variant comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:107, SEQ ID NO:108, or SEQ ID NO:

109.

111. The synthetic cytokine receptor according to any one of claims 2 to 110, wherein the variant IL-9R intracellular domain comprises the amino acid sequence of SEQ ID NO:107, SEQ ID NO:108, or SEQ ID NO:

109.

112. The synthetic cytokine receptor according to claim 3, wherein the chimeric JAK / STAT fusion domain comprises a JAK-binding domain derived from a type I cytokine receptor and a STAT-binding domain derived from an IL-9R intracellular domain.

113. The synthetic cytokine receptor according to claim 112, wherein the IL-9R STAT-binding domain comprises amino acid residues 73 to 230 of SEQ ID NO:

8.

114. The synthetic cytokine receptor according to claim 112 or claim 113, wherein the STAT-binding domain is 59 to 158 amino acids long and includes an IL-9R STAT-binding motif.

115. The synthetic cytokine receptor according to claim 114, wherein the IL-9R STAT binding motif comprises YLPQ (SEQ ID NO: 171).

116. The synthetic cytokine receptor according to any one of claims 112 to 115, wherein the IL-9R STAT binding domain is a truncated IL-9R STAT binding domain lacking a continuous amino acid sequence at the N-terminus of SEQ ID NO:

8.

117. The synthetic cytokine receptor according to any one of claims 112 to 116, wherein the IL-9R STAT binding domain is a truncated IL-9R STAT binding domain lacking a continuous amino acid sequence at the C-terminus of SEQ ID NO:

8.

118. The synthetic cytokine receptor according to any one of claims 112 to 117, wherein the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking amino acids at positions 1 to 72 and / or 132 to 230 of SEQ ID NO:

8.

119. The synthetic cytokine receptor according to any one of claims 112 to 118, wherein the IL-9R STAT-binding domain comprises an amino acid sequence that is at least about 85% identical to SEQ ID NO:122 or SEQ ID NO:

123.

120. The synthetic cytokine receptor according to any one of claims 112 to 119, wherein the IL-9R STAT binding domain comprises the amino acid sequence of SEQ ID NO:122 or SEQ ID NO:

123.

121. The synthetic cytokine receptor according to any one of claims 112 to 120, wherein the type I cytokine receptor is selected from the group consisting of interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R), interleukin-7 receptor (IL-7R), interleukin-13 receptor (IL-13R), interleukin-15 receptor (IL-15R), and interleukin-2 receptor (IL-21R).

122. The synthetic cytokine receptor according to any one of claims 112 to 121, wherein the type I cytokine receptor is IL-7R.

123. The synthetic cytokine receptor according to any one of claims 112 to 122, wherein the IL-7R JAK binding domain is 65 amino acids long and includes a box-1 motif.

124. The synthetic cytokine receptor according to any one of claims 112 to 123, wherein the IL-7R JAK binding domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

121.

125. The synthetic cytokine receptor according to any one of claims 112 to 124, wherein the IL-7R JAK binding domain comprises the amino acid sequence of SEQ ID NO:

121.

126. The synthetic cytokine receptor according to any one of claims 112 to 125, wherein the chimeric JAK / STAT fusion domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:114, SEQ ID NO:116, or SEQ ID NO:

181.

127. The synthetic cytokine receptor according to any one of claims 112 to 126, wherein the chimeric JAK / STAT fusion domain comprises the amino acid sequence of SEQ ID NO: 114, SEQ ID NO: 116, or SEQ ID NO:

181.

128. A synthetic cytokine receptor according to any one of claims 1 to 127, comprising an amino acid sequence that is at least about 85% identical to SEQ ID NO:113, SEQ ID NO:115, or SEQ ID NO:

117.

129. A synthetic cytokine receptor according to any one of claims 1 to 128, comprising the amino acid sequence of SEQ ID NO:113, SEQ ID NO:115, or SEQ ID NO:

117.

130. A synthetic cytokine receptor according to any one of claims 1 to 3 and 14 to 129, which is a constitutively active cytokine receptor.

131. A synthetic cytokine receptor according to any one of claims 1 to 130, which induces signal transduction via the STAT1 pathway, the STAT3 pathway, and / or the STAT5 pathway.

132. The synthetic cytokine receptor according to claim 131, wherein signaling via STAT1, STAT3, and / or STAT5 is increased compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R.

133. The synthetic cytokine receptor according to claim 131 or claim 132, wherein signaling via STAT1, STAT3, and / or STAT5 is sustained for a longer period compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R.

134. The synthetic cytokine receptor according to claim 133, wherein sustained STAT1 signaling, STAT3 signaling, and / or STAT5 signaling are determined by the phosphorylation state of STAT1, STAT3, and / or STAT5.

135. A polynucleotide encoding a synthetic cytokine receptor according to any one of claims 1 to 134.

136. A vector comprising a polynucleotide according to claim 135, wherein the vector is optionally a viral vector.

137. A method for modifying isolated cells, comprising the step of contacting the cells with a polynucleotide according to claim 135 or a vector according to claim 136.

138. Modified cells expressing the synthetic cytokine receptor according to any one of claims 1 to 137.

139. Modified cells expressing a synthetic cytokine receptor, which is a homodimer of the same polypeptide chain containing an extracellular domain, a transmembrane domain, and an intracellular domain capable of interleukin-9 receptor (IL-9R) signaling.

140. The modified cell according to claim 139, wherein the IL-9R signaling-capable intracellular domain comprises an IL-9R intracellular domain or a variant thereof.

141. The modified cell according to claim 139 or claim 140, wherein the intracellular domain capable of IL-9R signaling includes a chimeric JAK / STAT fusion domain.

142. Modified cells expressing a synthetic cytokine receptor, wherein the synthetic cytokine receptor comprises an extracellular domain, a transmembrane domain, and an intracellular domain of the interleukin-9 receptor (IL-9R) or a variant thereof, and the synthetic cytokine receptor is a constitutively active cytokine receptor.

143. The modified cell according to any one of claims 139 to 142, wherein the synthetic cytokine receptor is a multimer.

144. The modified cell according to any one of claims 139 to 142, wherein the synthetic cytokine receptor is a polymer of the same polypeptide chain comprising an extracellular domain, a transmembrane domain, and an IL-9R intracellular domain or a variant thereof.

145. The modified cell according to claim 143 or claim 144, wherein the polymer is a dimer.

146. The modified cell according to claim 145, wherein the dimer is a homodimer.

147. A modified cell according to any one of claims 138 to 146, wherein each polypeptide chain is constitutively polymerized.

148. The modified cell according to any one of claims 139 to 147, wherein the synthetic cytokine receptor comprises at least one self-assembling domain.

149. The modified cell according to claim 148, wherein the at least one self-assembling domain is the extracellular domain and / or the transmembrane domain.

150. The modified cell according to any one of claims 139 to 149, wherein the synthetic cytokine receptor is multimerized through the transmembrane domain and / or the extracellular domain.

151. The modified cell according to any one of claims 139 to 150, wherein the synthetic cytokine receptor is multimerized through the transmembrane domain and the extracellular domain.

152. Modified cells expressing a synthetic cytokine receptor, which is a homodimer of the same polypeptide chain containing an extracellular domain, a transmembrane domain, and an intracellular domain of the interleukin-9 receptor (IL-9R) or a variant thereof.

153. The modified cell according to any one of claims 139 to 152, wherein the extracellular domain and / or the transmembrane domain are heterogeneous with respect to IL-9R.

154. The modified cell according to any one of claims 139 to 153, wherein the transmembrane domain and the extracellular domain are transmembrane domain and extracellular domain derived from the same protein.

155. The modified cell according to any one of claims 139 to 153, wherein the transmembrane domain and extracellular domain are transmembrane domains and extracellular domains derived from different proteins.

156. The modified cell according to any one of claims 139 to 155, wherein the transmembrane domain has a length of 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, or 33 amino acids.

157. The modified cell according to any one of claims 139 to 156, wherein the transmembrane domain comprises a transmembrane domain derived from glycophorin A (GpA), carnitine palmitoyltransferase 1 (CPT1), tumor necrosis factor receptor (TNFR), or Muc24.

158. The modified cell according to any one of claims 139 to 156, wherein the transmembrane domain comprises a transmembrane domain derived from a thrombopoietin receptor.

159. The modified cell according to any one of claims 139 to 158, wherein the transmembrane domain promotes α-helix dimerization.

160. The modified cell according to any one of claims 139 to 159, wherein the transmembrane domain contains motif GXXXG (SEQ ID NO: 68).

161. The aforementioned transmembrane domain is a motif A modified cell according to any one of claims 139 to 159, including the modified cell.

162. The modified cell according to any one of claims 139 to 161, wherein the transmembrane domain is a transmembrane domain derived from glycophorin A (GpA), or a variant thereof containing one or more mutations (e.g., 1, 2, 3, 4, 5, or 6 mutations) compared to a wild-type GpA transmembrane domain, and the variant GpA is sufficient to promote α-helix dimerization.

163. The modified cell according to any one of claims 139 to 162, wherein the transmembrane domain comprises a transmembrane domain derived from glycophorin A (GpA).

164. The modified cell according to any one of claims 139 to 163, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:22 or SEQ ID NO:

43.

165. The modified cell according to any one of claims 139 to 164, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:22 or SEQ ID NO:

43.

166. The modified cell according to any one of claims 139 to 160, wherein the transmembrane domain comprises a GXXXG (SEQ ID NO: 68) motif and a GXXXA (SEQ ID NO: 69) motif.

167. The modified cell according to any one of claims 139-160 and 166, wherein the transmembrane domain comprises a transmembrane domain derived from carnitine palmitoyltransferase 1 (CPT1).

168. The modified cell according to any one of claims 139-160, 166, and 167, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:

45.

169. The modified cell according to any one of claims 139-160 and 166-168, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

45.

170. The modified cell according to any one of claims 139 to 169, wherein the synthetic cytokine receptor comprises motif AXXXA (SEQ ID NO: 77) or AXXXS (SEQ ID NO: 78).

171. The modified cell according to any one of claims 139 to 160 or 170, wherein the synthetic cytokine receptor comprises the motif ΦPXΦ (SEQ ID NO: 75) or ΦTXXAΦ (SEQ ID NO: 76).

172. The modified cell according to any one of claims 139-160, 170, or 171, wherein the transmembrane domain is derived from TNFR.

173. The modified cell according to claim 172, wherein the TNFR is TACI, DR5, p75NTR, Fas, TNFR1, TNFR2, or OX40.

174. The modified cell according to any one of claims 139-160 and 170-173, wherein the transmembrane domain comprises a transmembrane domain derived from DR5.

175. The modified cell according to any one of claims 139-160 and 170-174, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

46.

176. The modified cell according to any one of claims 139-160 and 170-175, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

46.

177. The modified cell according to any one of claims 139-160 and 170-174, wherein the transmembrane domain comprises a transmembrane domain derived from TACI.

178. The modified cell according to any one of claims 139-160, 170-174, and 177, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:

44.

179. A modified cell according to any one of claims 39 to 161, 170 to 174, 177, and 178, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

44.

180. The modified cell according to any one of claims 139 to 158, wherein the transmembrane domain comprises 1 to 6 cysteine ​​residues.

181. The modified cell according to any one of claims 139-158 and 180, wherein the transmembrane domain promotes disulfide bond dimerization.

182. The modified cell according to claim 181, wherein the disulfide bond dimerization forms 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

183. The modified cell according to any one of claims 139-158 and 180-182, wherein the transmembrane domain is a variant transmembrane domain containing one or more mutations compared to the wild-type transmembrane domain for promoting homodimerization of the receptor polypeptide.

184. The modified cell according to claim 183, wherein the one or more mutations promote α-helix dimerization or disulfide bond dimerization.

185. The modified cell according to claim 183 or claim 184, wherein the one or more mutations introduce at least one cysteine ​​into the transmembrane domain.

186. The modified cell according to any one of claims 183 to 185, wherein the one or more mutations introduce proline into the transmembrane domain.

187. The modified cell according to any one of claims 183 to 185, wherein the one or more mutations introduce threonine into the transmembrane domain.

188. The modified cell according to any one of claims 183 to 187, wherein the one or more mutations introduce a trimer peptide of cysteine, proline, and another amino acid other than cysteine ​​and proline into the transmembrane domain.

189. The modified cell according to any one of claims 183 to 188, wherein the one or more mutations introduce a cysteine-proline-threonine trimer peptide (CPT or TCP) into the transmembrane domain.

190. The transmembrane domain is a variant IL-7R transmembrane domain, and the one or more mutations are wild-type transmembrane sequences. A modified cell according to any one of claims 183 to 188, contained within.

191. The modified cell according to any one of claims 139-158 and 180-190, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:

21.

192. The modified cell according to any one of claims 139-158 and 180-191, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:

21.

193. The modified cell according to any one of claims 139 to 158, wherein the transmembrane domain comprises a transmembrane domain derived from Muc24.

194. The modified cell according to any one of claims 139-158 and 193, wherein the transmembrane domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

23.

195. The modified cell according to any one of claims 139-158, 193, or 194, wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO:

23.

196. The modified cell according to any one of claims 139 to 195, wherein the extracellular domain is approximately 150 to 260 amino acids long.

197. The modified cell according to any one of claims 139 to 196, wherein the extracellular domain is a dimerizing domain.

198. The modified cell according to claim 197, wherein the dimerization domain includes a hinge region.

199. The modified cell according to any one of claims 139 to 198, wherein the extracellular domain promotes disulfide bond dimerization.

200. The modified cell according to any one of claims 139 to 199, wherein the extracellular domain comprises 1 to 6 cysteine ​​residues.

201. The modified cell according to claim 199 or claim 200, wherein the disulfide bond dimerization forms 1 to 4 disulfide crosslinks between the polypeptide chains of the synthetic cytokine receptor.

202. The modified cell according to any one of claims 139 to 201, wherein the extracellular domain is derived from the extracellular domain of CD34, DAP12, glycophorin A, CD8, or Muc24.

203. The modified cell according to any one of claims 139 to 202, wherein the extracellular domain comprises an extracellular domain derived from a thrombopoietin receptor.

204. The modified cell according to any one of claims 139 to 203, wherein the extracellular domain comprises the extracellular domain of CD8, or a truncated portion thereof containing at least one cysteine ​​residue.

205. The modified cell according to any one of claims 139 to 204, wherein the extracellular domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:18 or SEQ ID NO:

19.

206. The modified cell according to any one of claims 139 to 205, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:18 or SEQ ID NO:

19.

207. The modified cell according to any one of claims 139 to 203, wherein the extracellular domain comprises the extracellular domain of CD34, or a truncated portion thereof containing at least one cysteine ​​residue.

208. The modified cell according to any one of claims 139-203 and 207, wherein the extracellular domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:

4.

209. The modified cell according to any one of claims 139-203, 207, and 208, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:

4.

210. The modified cell according to any one of claims 139 to 203, wherein the extracellular domain is the extracellular domain of Muc24, or a truncated portion thereof containing at least one cysteine ​​residue.

211. The modified cell according to any one of claims 139-203 and 210, wherein the extracellular domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

20.

212. The modified cell according to any one of claims 139-203, 210, and 211, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:

20.

213. The synthetic cytokine receptor according to any one of claims 139 to 203, wherein the extracellular domain is the extracellular domain of DAP12, or a truncated portion thereof containing at least one cysteine ​​residue.

214. The modified cell according to any one of claims 139-203 and 213, wherein the extracellular domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:

5.

215. The modified cell according to any one of claims 139-203, 213, and 214, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:

5.

216. The synthetic cytokine receptor according to any one of claims 139 to 203, wherein the extracellular domain is the extracellular domain of glycophorin A (GpA), or a truncated portion thereof containing at least one cysteine ​​residue.

217. The modified cell according to any one of claims 139-203 and 216, wherein the extracellular domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:16 or SEQ ID NO:

17.

218. The modified cell according to any one of claims 139-203, 216, and 217, wherein the extracellular domain comprises the amino acid sequence of SEQ ID NO:16 or SEQ ID NO:

17.

219. The modified cell according to any one of claims 139 to 218, wherein the IL-9R intracellular domain or a variant thereof is approximately 100 to 260 amino acids long.

220. The modified cell according to any one of claims 139 to 219, wherein the IL-9R intracellular domain or a variant thereof comprises a box 1 motif and / or a box 2 motif.

221. The modified cell according to any one of claims 139 to 220, wherein the IL-9R intracellular domain or a variant thereof includes a box 2 motif.

222. The modified cell according to any one of claims 2 and 4 to 80, wherein the IL-9R intracellular domain or a variant thereof is 230 amino acid long.

223. The modified cell according to any one of claims 139 to 222, wherein the IL-9R intracellular domain or its variant is the wild-type IL-9R intracellular domain, or a variant thereof containing one or more mutations compared to the wild-type IL-9R intracellular domain shown in SEQ ID NO:

8.

224. The modified cell according to any one of claims 139 to 223, wherein the one or more mutations include the insertion, deletion, and / or substitution of one or more amino acids.

225. The modified cell according to claim 223 or claim 224, wherein the one or more mutations promote signaling through the STAT1 pathway, the STAT3 pathway, and / or the STAT5 pathway.

226. The genetically modified cell according to any one of claims 139 to 225, wherein the IL-9R intracellular domain or a variant thereof contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:

8.

227. The modified cell according to any one of claims 139 to 226, wherein the IL-9R intracellular domain or a variant thereof comprises the amino acid sequence of SEQ ID NO:

8.

228. The modified cell according to any one of claims 139 to 225, wherein the IL-9R intracellular domain or a variant thereof contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, or SEQ ID NO:

56.

229. The modified cell according to any one of claims 139 to 225 and 228, wherein the IL-9R intracellular domain or its variant comprises the amino acid sequence of SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, or SEQ ID NO:

56.

230. The modified cell according to any one of claims 139 to 229, wherein the synthetic cytokine receptor contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:

67.

231. The modified cell according to any one of claims 139 to 230, wherein the synthetic cytokine receptor comprises the amino acid sequence of SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:66, or SEQ ID NO:

67.

232. The modified cell according to any one of claims 140 to 231, wherein the IL-9R intracellular domain or its variant comprises one or more amino acid deletions relative to the wild-type IL-9R intracellular domain (SEQ ID NO: 8).

233. The modified cell according to any one of claims 140 to 232, wherein the IL-9R intracellular domain or its variant is a truncated IL-9R lacking a continuous amino acid sequence at the C-terminus of the wild-type IL-9R intracellular domain.

234. The modified cell according to claim 233, wherein the shortened IL-9R intracellular domain or its variant is shortened by 62 to 99 consecutive amino acids from the C-terminus of the wild-type IL-9R intracellular domain.

235. The modified cell according to any one of claims 140 to 232, wherein the IL-9R intracellular domain or a variant thereof is a shortened IL-9R lacking amino acids 132 to 230 of SEQ ID NO:8 or lacking amino acids 134 to 230 of SEQ ID NO:

8.

236. The modified cell according to any one of claims 140 to 235, wherein the IL-9R intracellular domain or a variant thereof contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:54, SEQ ID NO:103, or SEQ ID NO:

104.

237. The modified cell according to any one of claims 140 to 236, wherein the IL-9R intracellular domain or a variant thereof comprises the amino acid sequence of SEQ ID NO: 54, SEQ ID NO: 103, or SEQ ID NO:

104.

238. The modified cell according to any one of claims 140 to 237, wherein the synthetic cytokine receptor contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, or SEQ ID NO:

63.

239. The modified cell according to any one of claims 140 to 238, wherein the synthetic cytokine receptor comprises the amino acid sequence of SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, or SEQ ID NO:

63.

240. The genetically modified cell according to any one of claims 140 to 225, wherein the IL-9R intracellular domain or its variant comprises one or more amino acid substitutions relative to the wild-type IL-9R intracellular domain (SEQ ID NO: 8).

241. The modified cell according to claim 240, wherein the IL-9R intracellular domain or a variant thereof comprises a STAT-binding motif or a variant thereof.

242. The modified cell according to claim 241, wherein the STAT binding motif comprises a STAT1 binding motif, a STAT3 binding motif, and / or a STAT5 binding motif.

243. The modified cell according to claim 241 or claim 242, wherein the STAT-binding motif includes YLPQ (SEQ ID NO: 171).

244. The modified cell according to any one of claims 241 to 243, wherein the STAT-binding motif includes a variant STAT-binding motif.

245. The modified cell according to any one of claims 241 to 244, wherein the variant STAT-binding motif includes YRPQ (SEQ ID NO: 172).

246. The modified cell according to any one of claims 241 to 244, wherein the variant STAT-binding motif includes YLPL (SEQ ID NO: 173).

247. The modified cell according to any one of claims 241 to 244, wherein the variant STAT-binding motif includes YLKQ (SEQ ID NO: 174).

248. The modified cell according to any one of claims 140 to 247, wherein the variant IL-9R intracellular domain or the variant contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:107, SEQ ID NO:108, or SEQ ID NO:

109.

249. The modified cell according to any one of claims 140 to 248, wherein the variant IL-9R intracellular domain comprises the amino acid sequence of SEQ ID NO:107, SEQ ID NO:108, or SEQ ID NO:

109.

250. The modified cell according to claim 141, wherein the chimeric JAK / STAT fusion domain comprises a JAK-binding domain derived from a type I cytokine receptor and a STAT-binding domain derived from an IL-9R intracellular domain.

251. The modified cell according to claim 250, wherein the IL-9R STAT-binding domain comprises amino acid residues 73-230 of SEQ ID NO:

8.

252. The modified cell according to claim 250 or claim 251, wherein the STAT-binding domain is 59 to 158 amino acids long and contains an IL-9R STAT-binding motif.

253. The modified cell according to claim 252, wherein the IL-9R STAT binding motif includes YLPQ (SEQ ID NO: 171).

254. The modified cell according to any one of claims 250 to 253, wherein the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking a continuous amino acid sequence at the N-terminus of SEQ ID NO:

8.

255. The modified cell according to any one of claims 250 to 254, wherein the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking a continuous amino acid sequence at the C-terminus of SEQ ID NO:

8.

256. The modified cell according to any one of claims 250 to 255, wherein the IL-9R STAT-binding domain is a shortened IL-9R STAT-binding domain lacking amino acids at positions 1 to 72 and / or 132 to 230 of SEQ ID NO:

8.

257. The modified cell according to any one of claims 250 to 256, wherein the IL-9R STAT-binding domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:122 or SEQ ID NO:

123.

258. The modified cell according to any one of claims 250 to 257, wherein the IL-9R STAT binding domain comprises the amino acid sequence of SEQ ID NO:122 or SEQ ID NO:

123.

259. The modified cell according to any one of claims 250 to 258, wherein the type I cytokine receptor is selected from the group consisting of interleukin-2 receptor (IL-2R), interleukin-4 receptor (IL-4R), interleukin-7 receptor (IL-7R), interleukin-13 receptor (IL-13R), interleukin-15 receptor (IL-15R), and interleukin-2 receptor (IL-21R).

260. The modified cell according to any one of claims 250 to 259, wherein the type I cytokine receptor is IL-7R.

261. A modified cell according to any one of claims 250 to 260, wherein the IL-7R JAK binding domain is 65 amino acids long and contains a box-1 motif.

262. The modified cell according to any one of claims 250 to 261, wherein the IL-7R JAK binding domain contains an amino acid sequence that is at least about 85% identical to that of SEQ ID NO:

121.

263. The modified cell according to any one of claims 250 to 262, wherein the IL-7R JAK binding domain comprises the amino acid sequence of SEQ ID NO:

121.

264. The modified cell according to any one of claims 250 to 263, wherein the chimeric JAK / STAT fusion domain contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:114, SEQ ID NO:116, or SEQ ID NO:

181.

265. The modified cell according to any one of claims 250 to 264, wherein the chimeric JAK / STAT fusion domain comprises the amino acid sequence of SEQ ID NO:114, SEQ ID NO:116, or SEQ ID NO:

181.

266. The modified cell according to any one of claims 139 to 265, wherein the synthetic cytokine receptor contains an amino acid sequence that is at least about 85% identical to SEQ ID NO:113, SEQ ID NO:115, or SEQ ID NO:

117.

267. The modified cell according to any one of claims 139 to 266, wherein the synthetic cytokine receptor comprises the amino acid sequence of SEQ ID NO:113, SEQ ID NO:115, or SEQ ID NO:

117.

268. The modified cell according to any one of claims 139-141 and 152-267, wherein the synthetic cytokine receptor is a constitutively active cytokine receptor.

269. The modified cell according to any one of claims 139 to 268, wherein the synthetic cytokine receptor induces signal transduction through the STAT1 pathway, the STAT3 pathway, and / or the STAT5 pathway.

270. The modified cell according to claim 269, wherein signaling through STAT1, STAT3, and / or STAT5 is increased compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R.

271. Modified cells according to claim 269 or claim 270, wherein signaling via STAT1, STAT3, and / or STAT5 is sustained for a longer period compared to STAT1, STAT3, and / or STAT5 signaling via wild-type IL-9R.

272. The modified cell according to claim 271, wherein sustained STAT1 signaling, STAT3 signaling, and / or STAT5 signaling are determined by the phosphorylation state of STAT1, STAT3, and / or STAT5.

273. A modified cell according to any one of claims 139 to 272, further expressing at least one different type of modified receptor.

274. The modified cell according to claim 273, wherein the at least one different type of modified receptor is a chimeric antigen receptor.

275. The modified cell according to claim 274, wherein the extracellular domain of the chimeric antigen receptor binds to an antigen expressed in cancer cells.

276. The modified cell according to claim 275, wherein the extracellular domain of the chimeric antigen receptor binds to the antibody idiotype.

277. The modified cell according to claim 276, wherein the antibody is against an antigen expressed in cancer cells.

278. The modified cell according to claim 277, wherein the cancer cells are blood cancer cells or solid tumor cancer cells.

279. The modified cell according to any one of claims 139 to 278, wherein the cell is an immune cell.

280. The modified cell according to any one of claims 139 to 279, wherein the cell is a lymphocyte.

281. A modified cell according to any one of claims 139 to 280, which is an immune effector cell.

282. The modified cell according to any one of claims 139 to 281, wherein the cell is a T cell or a natural killer (NK) cell.

283. The modified cell according to any one of claims 139 to 282, wherein the cell is a T cell, and the T cell is a CD4+ T cell or a CD8+ T cell.

284. The modified cell according to claim 283, wherein the immune effector cell is a cytotoxic T cell.

285. The modified cell according to claim 283, wherein the immune effector cells are natural killer cells.

286. The modified cell according to any one of claims 139 to 285, wherein the cell is a primary cell.

287. The modified cell according to any one of claims 139 to 286, wherein the cell is a human cell.

288. A cell population comprising at least one modified cell according to any one of claims 139 to 287.

289. The cell population according to claim 288, wherein the at least one modified cell comprises a modified CD4+ T cell and a modified CD8+ T cell.

290. A pharmaceutical composition comprising modified cells according to any one of claims 90 to 191 or a cell population according to claim 288 or claim 289.

291. The pharmaceutical composition according to claim 290, further comprising a pharmaceutically acceptable carrier.

292. The pharmaceutical composition according to claim 290 or claim 291, further comprising a cryoprotective substance.

293. A pharmaceutical composition according to any one of claims 290 to 292, for use in the treatment of cancer in a subject.

294. A method for treating a disease or condition in a subject, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition according to any one of claims 290 to 293 to the subject.

295. The method according to claim 294, wherein the disease or condition is cancer.

296. The method according to claim 294 or claim 295, wherein the modified cells of the pharmaceutical composition express a modified antigen receptor that binds to an antigen expressed in the cancer cells.

297. The method according to claim 296, wherein the modified antigen receptor is a chimeric antigen receptor.

298. The method according to claim 296, wherein the modified antigen receptor is a T cell receptor.

299. A method for improving the function of immune cells, comprising the step of introducing a polynucleotide according to claim 135 or a vector according to claim 136 into immune cells, wherein the improvement in immune cell function includes an increase in STAT1 signaling, STAT3 signaling, and / or STAT5 signaling compared to immune cells expressing wild-type IL-9R.

300. A method for improving the cytotoxicity of immune cells, comprising the step of introducing a polynucleotide according to claim 135 or a vector according to claim 136 into immune cells, wherein the improvement in the cytotoxicity of immune cells includes an increase in target cell killing compared to immune cells expressing wild-type IL-9R.

301. A method for improving the viability of immune cells, comprising the step of introducing a polynucleotide according to claim 135 or a vector according to claim 136 into immune cells, wherein the improvement in the viability of immune cells includes a reduction in immune cell death compared to immune cells expressing wild-type IL-9R.

302. A method for improving cytokine secretion by immune cells, comprising the step of introducing a polynucleotide according to claim 135 or a vector according to claim 136 into immune cells, wherein the improvement in cytokine secretion includes an increase in interferon secretion compared to immune cells expressing wild-type IL-9R.

303. The method according to claim 302, wherein the interferon comprises IFN-γ.

304. The method according to any one of claims 299 to 303, wherein the immune cells further comprise a modified antigen receptor.

305. The method according to claim 304, wherein the one different type of modified receptor is a chimeric antigen receptor (CAR).

306. The method according to claim 305, wherein the extracellular domain of the CAR binds to an antigen expressed in cancer cells.

307. The method according to claim 306, wherein the cancer cells are blood cancer cells or solid tumor cancer cells.

308. The method according to any one of claims 299 to 307, wherein the immune cells are lymphocytes.

309. The method according to any one of claims 299 to 308, wherein the immune cells are effector cells.

310. The method according to any one of claims 299 to 309, wherein the immune cells are T cells or NK cells.

311. The method according to any one of claims 299 to 303, wherein the immune cells are T cells.

312. The method according to any one of claims 299 to 310, wherein the immune cell is a T cell, and the T cell is a CD4+ T cell or a CD8+ T cell.

313. The method according to any one of claims 299 to 310, wherein the immune cells are cytotoxic T cells.

314. The method according to any one of claims 299 to 311, wherein the immune cells are NK cells.

315. The method according to any one of claims 299 to 313, wherein the immune cells are primary cells.

316. A method for improving the function of immune cells, comprising the step of introducing a polynucleotide according to claim 135 or a vector according to claim 136 into immune cells, thereby improving the function of said immune cells.

317. The method according to claim 316, wherein improving immune cell function includes one or more of the following compared to reference cells: increased STAT signaling, increased cytotoxicity, increased proliferation, increased viability, increased cytokine secretion, and increased cytotoxic protein secretion.

318. The method according to claim 317, wherein the reference cells include unmodified immune cells or modified immune cells.

319. The method according to claim 318, wherein the modified immune cells express a modified antigen receptor.

320. The method according to claim 319, wherein the modified antigen receptor is wild-type IL-9R, a chimeric antigen receptor (CAR), or a T cell receptor (TCR).

321. The method according to any one of claims 317 to 320, wherein the increase in STAT signaling includes an increase in STAT1 signaling, STAT3 signaling, and / or STAT5 signaling.

322. The method according to any one of claims 317 to 321, wherein the increased cytotoxicity includes increased killing of target cells.

323. The method according to any one of claims 317 to 322, wherein the increase in cytokine secretion includes an increase in the secretion of one or more of interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-10 (IL-10), and TNFα.

324. The method according to any one of claims 317 to 323, wherein the increase in cytotoxic protein secretion includes an increase in the secretion of one or more of granzyme A, granzyme B, granulosin, and perforin.

325. The method according to any one of claims 317 to 324, wherein the increase in viability includes a reduction in cell death.

326. The method according to any one of claims 316 to 325, wherein the immune cells are lymphocytes.

327. The method according to any one of claims 316 to 326, wherein the immune cells are effector cells.

328. The method according to any one of claims 316 to 327, wherein the immune cells are T cells or NK cells.

329. The method according to any one of claims 316 to 327, wherein the immune cell is a T cell, and the T cell is a CD4+ T cell or a CD8+ T cell.

330. The method according to any one of claims 316 to 329, wherein the immune cells are cytotoxic T cells.

331. The method according to any one of claims 316 to 328, wherein the immune cells are NK cells.

332. The method according to any one of claims 316 to 331, wherein the immune cells are primary cells.

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