Activation of t-cells with mutant g-csfr

Abstract

Provided herein are particles or compositions comprising one or more nucleic acid molecules encoding a chimeric antigen receptor (CAR) and / or a polypeptide comprising i) a ligand binding extracellular domain (ECD), ii) an immune cell activating intracellular domain (ICD); and iii) transmembrane domains linking the ECD and the ICD, polypeptides comprising these domains and nucleic acid molecules encoding these polypeptides. Also provided herein are compositions comprising these nucleic acid molecules and methods of using these compositions. Also provided herein are viral particles comprising these nucleic acid molecules.

Description

(19) State Intellectual Property Office (12) Invention Patent Application (10) Application Publication Number (43) Application Publication Date (21) Application Number 202480046363.0 (22) Application Date 2024.06.13 (30) Priority Data 63 / 508,055 2023.06.14 US 63 / 510,745 2023.06.28 US (85) PCT International Application Entering National Phase Date 2026.01.09 (86) PCT International Application Application Data PCT / US2024 / 033821 2024.06.13 (87) PCT International Application Publication Data WO2024 / 259109 EN 2024.12.19 (71) Applicant: Interus Biotherapeutics, Inc. Address: Pennsylvania, USA (72) Inventors: S.J. Doolittle, P.R. Johnson (74) Patent Agency: Beijing Liu Shen Law Firm, 11105 Patent Attorney: Qin Jian (51) Int.Cl. C07K 14 / 705 (2006.01) C12N 15 / 86 (2006.01) (54) Invention Title: Activation of T Cells with Mutant G-CSFR (57) Abstract This document provides particles or compositions comprising one or more nucleic acid molecules encoding a chimeric antigen receptor (CAR) and / or a polypeptide comprising i) a ligand-binding extracellular domain (ECD), ii) an immune cell activation intracellular domain (ICD); and iii) a transmembrane domain linking the ECD and the ICD, a polypeptide comprising these domains, and a nucleic acid molecule encoding the polypeptide. This document also provides compositions comprising these nucleic acid molecules and methods of using these compositions. This document also provides viral particles comprising these nucleic acid molecules. Claims (6 pages), Description (166 pages), Sequence List (electronic publication), Drawings (78 pages), CN 121487962 A, 2026.02.06, CN 1 21 48 79 62 A

Claims

1. A particle or composition comprising one or more nucleic acid molecules encoding: a) a chimeric antigen receptor (CAR); and b) a polypeptide comprising: i) a ligand-binding extracellular domain (ECD); ii) an immune cell activating intracellular domain (ICD); and iii) a transmembrane domain linking the ECD to the ICD.

2. The particle of claim 1, wherein the immune cell activating ICD comprises an ICD of hGCSF-R or a mutant or fragment thereof.

3. The particle of claim 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111, or an amino acid sequence having at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity thereto.

4. The particle of claim 3, wherein the hGCSF-R ICD does not comprise the sequence of SEQ ID NO:

110.

5. The particle of claim 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111 and does not comprise amino acid residues 800-813, 775-813, 750-813, 725-813, or 716-813 of SEQ ID NO: 125, or a fragment thereof located at the C-terminal residues of SEQ ID NO:

111.

6. The particle of claim 2, wherein the hGCSF-R ICD comprises the sequence of SEQ ID NO: 111 and amino acid residues 716-812 of SEQ ID NO: 125, or a fragment thereof located at the C-terminal residues of SEQ ID NO: 111, provided that the ICD does not comprise a contiguous fragment of amino acid residues 716-813 of SEQ ID NO:

125.

7. The particle of claim 1, wherein the ECD is a GCSF-R ECD, optionally wherein the GCSF-R ECD is a human GCSF-R ECD comprising the amino acid sequence of SEQ ID NO: 107 or SEQ ID NO:

108.

8. The particle of claim 1, wherein the ECD comprises an ECD of human growth hormone receptor (hGH-R) or a mutant or fragment thereof, wherein the ECD of human growth hormone receptor (hGH-R) comprises an amino acid sequence encoded by exons 1-7 of hGH-R or a mutant or fragment thereof.

9. The particle of claim 8, wherein the ECD of hGH-R comprises a mutation of C259R.

10. The particle of claim 8, wherein the ECD of the hGH-R comprises the amino acid sequence of SEQ ID NO: 114 or SEQ ID NO: 115, or an amino acid sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity thereto.

11. The particle of claim 1, wherein the ICD comprises an ICD of glycoprotein 130 ("gpl30 ICD") or a variant or fragment thereof.

12. The particle of claim 1, wherein the ICD comprises a fragment of the GCSF-R ICD, wherein the GCSF-R ICD fragment does not comprise a STAT3, SH3, and / or SHC domain.

13. The particle of claim 1, wherein the ICD comprises an IL-2Rβ ICD or a fragment or variant thereof.

14. The particle of claim 1, wherein the ICD comprises an IL-7Rα ICD or a fragment or variant thereof.

15. The particle of claim 1, wherein the ICD comprises a gpl30 ICD or a variant or fragment thereof linked to an IL-2Rβ ICD or a fragment thereof.

16. The particle of claim 1, wherein the transmembrane domain comprises a GCSF-R, gpl30, or hGH-R transmembrane domain.

17. The particle of claim 1, wherein the polypeptide comprises: a) an ECD that is a GCSFR-ECD or a variant thereof; a transmembrane domain that is a GCSF-R transmembrane domain or a variant thereof; and an immunostimulatory ICD that is a GCSF-R ICD or a variant thereof; b) an ECD that is a hGH-R-ECD or a variant thereof; a transmembrane domain that is a GCSF-R transmembrane domain or a variant thereof; and an immunostimulatory ICD that is a GCSF-R ICD or a variant thereof; c) an ECD that is a hGH-R-ECD or a variant thereof; a transmembrane domain that is a gpl30 transmembrane domain or a variant thereof; and an immunostimulatory ICD comprising a gpl30 ICD or a variant thereof and an IL-2Rβ ICD or a variant thereof; or d) an ECD that is a hGH-R-ECD or a variant thereof; a transmembrane domain that is a GCSF-R transmembrane domain or a variant thereof; and an immunostimulatory ICD comprising a GCSF-R ICD or a variant thereof and an IL-7Rα ICD or a variant thereof.

18. The particle of claim 1, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124, or a variant of any of the foregoing. ​ ​ ​ ​ 19. The particle of claim 18, wherein the variant comprises a polypeptide that is at least 70%, 71%, 72%, 73%, 74%, 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: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO:

124.

20. The particle of claim 1, wherein the CAR comprises an antigen binding domain that binds to CD20.

21. The particle of claim 20, wherein the CAR comprises an amino acid sequence that is at least 85% identical to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO: 129, at least 90% identical to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO: 129, at least 95% identical to SEQ ID NO: 99, at least 99% identical to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO: 129, or at least 100% identical to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO:

129.

22. The particle of any one of claims 1-21, wherein the particle is a vector, a virus, a liposome, a plasmid, or a cell.

23. The particle of any one of claims 1-22, wherein the particle is a viral vector, wherein the viral vector is a lentiviral vector, an AV vector, an AAV vector, or the like.

24. A nucleic acid molecule encoding a polypeptide, the nucleic acid molecule comprising an isolated nucleic acid molecule, the polypeptide comprising: i) a ligand-binding extracellular domain (ECD); ii) an immune cell activating intracellular domain (ICD); and iii) a transmembrane domain linking the ECD to the ICD.

25. The nucleic acid molecule of claim 24, wherein the nucleic acid molecule encodes a polypeptide of any one of claims 1-19.

26. The nucleic acid molecule of claim 24, wherein the polypeptide comprises an amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124, or a variant of any of the foregoing.

27. The nucleic acid molecule of claim 26, wherein the variant comprises a polypeptide having at least 70%, 71%, 72%, 73%, 74%, 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% identity to SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 16, or SEQ ID NO:

20.

28. The nucleic acid molecule of any one of claims 24-27, wherein the nucleic acid molecule further encodes a chimeric antigen receptor (CAR).

29. A viral particle comprising a heterologous viral glycoprotein, a targeting moiety, and one or more nucleic acid molecules encoding one or more heterologous molecules of interest, wherein the targeting moiety comprises a polypeptide having the formula T-S1, wherein T is a target binding domain and S1 is a stalk moiety, and wherein the nucleic acid molecule encoding a heterologous molecule of interest is a nucleic acid molecule of any one of claims 24-28.

30. The viral particle of claim 29, wherein the target binding domain (T) binds to: CD7, CD8, cKit (CD117), CD4, CD3, CD5, CD6, CD2, TCR a, TCR β, TCR γ, TCR δ, CD10, CD34, CD110, CD33, CD14, CD68, CCR7, CD62L, CD25, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, or CXCR3, a glycosylated CD43 epitope expressed on acute leukemias or lymphomas but not on hematopoietic progenitor cells; a glycosylated CD43 epitope expressed on non-hematopoietic cancers; A kinase anchor protein 4 (AKAP-4); adrenergic receptor beta 3 (ADRB3); AFP; anaplastic lymphoma kinase (ALK); androgen receptor; angiopoietin-binding cell surface receptor 2 (Tie 2); anti-desmoglein 1 (Dsgl) autoantibody; anti-desmoglein 3 (Dsg3) autoantibody; B7H3 (CD276); biotin; bone marrow stromal cell antigen 2 (BST2); BST1 / CD157; cancer / testis antigen 1 (NY-ESO-1); cancer / testis antigen 2 (LAGE-la); carbonic anhydrase IX (CAIX); carcinoembryonic antigen (CEA); CCCTC-binding factor (Zinc Finger Protein)-like (BORIS or Brother of the Regulator of Imprinted Sites); CCR4; CD5; CD19; CD20; CD22; CD24; CD30; CD32 (FCGR2A); CD33; CD34; CD38; CD44v6; CD72; CD79a; CD79b; CD97; CD99; CD123; CD171; CD179a; CD179b-IGLll; CD200R; CD276 / B7H3; CD300 molecule-like family member f (CD300LF); CDH1-CD324; CDH6; CDH17; CDH19; Chromosome X Open Reading Frame 61 (CXORF61); Claudin 6 (CLDN6); Claudin 18 8.2 (CLD18A2 or CLDN18A.2); CMV pp65; C-MYC epitope tag; Cripto; CS1 (also known as CD2 subset 1 or CRACC or SLAMF7 or CD319 or 19A24); CSF2RA (GM-CSFR-alpha); C-type lectin domain family 12 member A (CLEC12A); C-type lectin-like molecule-1 (CLL-1 or CLECL1); Cyclin Bl; Cytochrome P450 IB 1 (CYP1B 1); DLL3; EBV-EBNA3c; EGF-bike module 2 containing mucin-like hormone receptor-like (EMR2); Elongation factor 2 mutant (ELF2M); Ephrin B2; Ephrin type-A receptor 2 (EphA2); Epidermal growth factor receptor (EGFR); Epidermal growth factor receptor variant III (EGFRviii); Epithelial cell adhesion molecule (EPCAM); ERG; ETS translocation-variant gene 6 located on chromosome 12p (ETV6-AML); Fc fragment of IgA receptor (FCAR or CD89); Fc receptor like 5 (FCRL5); Fibroblast activation protein alpha (FAP); FITC; Fms-like tyrosine kinase 3 (FLT3); Folate receptor alpha (Fra or FR1); Folate receptor beta (FRb); Follicle-stimulating hormone receptor (FSHR); Fos-related antigen 1; Fucosyl-GMl; G protein-coupled receptor class C group 5 member D (GPRC5D); G protein-coupled receptor 20 (GPR20); GAD; Ganglioside G2 (GD2); Ganglioside GD3 (aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(l-4)bDGlcp(l-l)Cer); Ganglioside GM3 (aNeu5Ac(2-3)bDClalp(l-4)bDGlcp(l-l)Cer); GD3; GFR alpha 4; Glycoprotein 100 (gplOO); Glypican-3 (GPC3); Gonadotropin receptor (CGHR or GR); GpA33; GpNMB; GPRC5D; Guanylate cyclase C (GCC); Mutated heat shock protein 70-2 (mut hsp70-2); Hepatitis A virus cellular receptor 1 (HAVCR1); Hexose portion of globoH glycosphingolipid (GloboH); High molecular weight-melanoma-associated antigen (HMWMAA); HIV1 envelope glycoprotein; HLA; HLA-DOA; HLA-A; HLA-A2; HLA-B; HLA-C; HLA-DM; HLA-DOB; HLA-DP; HLA-DQ; HLA-DR; HLA-G;. HTLVl-Tax; human papilloma virus E6 (HPV E6); human papilloma virus E7 (HPV E7); human telomerase reverse transcriptase (hTERT); IgE; IL13Ra2; Ill lRa; immunoglobulin lambda-like polypeptide 1 (IGLL1); influenza A hemagglutinin (HA); insulin-like growth factor 1 receptor (IGF-I receptor); interleukin 11 receptor alpha (IL-llRa); interleukin 13 receptor subunit alpha-2 (IL-13Ra2 or CD213A2); intestinal carboxyl esterase; KIT (CD117); KSHV K8.1; KSHV-gH; LAMP1; legumain; leukocyte immunoglobulin-like receptor subfamily A member 2 (LILRA2); leukocyte-associated immunoglobulin-like receptor 1 (LAIR1); luteinizing hormone receptor (LHR); Lewis (Y) antigen; Lews Ag; Livl; locus K 9 (LY6K); low conductance chloride ion channel; lymphocyte antigen 6 complex; lymphocyte antigen 75 (LY75); lymphocyte-specific protein tyrosine kinase (LCK); mammary differentiation antigen (NY-BR-1); melanoma antigen recognized by T cells 1 (MelanA or MARTI); melanoma-associated antigen 1 (MAGE-A1); melanoma cancer testis antigen-1 (MAD-CT-1); melanoma cancer testis antigen-2 (MAD-CT-2); melanoma cell apoptosis inhibitor (ML-IAP); mesothelin; MPL; mucin 1, cell surface associated (MUC1); N-acetylglucosaminyltransferase V (NA17); nectin-4; neural cell adhesion molecule (NCAM); NKG2D; NYBR1; O-acety-GD2 ganglioside (OacGD2); olfactory receptor 51E2 (OR51E2); oncogenic fusion protein consisting of breakpoint cluster region (BCR) and Abelson murine leukemia viral oncogene homolog 1 (Abl) (bcr-abl); P53 mutant; paired box protein Pax-3 (PAX3); Pax-box protein Pax-5 (PAX5); pan adaptor protein 3 (PANX3); PDL1; P-glycoprotein; placenta-specific 1 (PLAC1); platelet-derived growth factor receptor beta (PDGFR-beta); polysialic acid; proacrosomal antigen sp32 (OY-TES1); prostein; prostate cancer tumor antigen-1 (PCT A-1 or galectin 8); prostate stem cell antigen (PSCA); prostate-specific membrane antigen (PSMA); prostatic acid phosphatase (PAP); prostein; serine protease 21 (Testisin or PRSS21); proteasome (Prosome Macropain) subunit beta type 9 (LMP2); PTK7; Ras G12V; Ras homolog family member C (RhoC); rat sarcoma (Ras) mutant; receptor for advanced glycation end products (RAGE-1); receptor tyrosine kinase-like orphan receptor 1 (ROR1); receptor tyrosine-protein kinase ERBB2 or Her-22 / neu; renal ubiquitous protein 1 (RU1); renal ubiquitous protein 2 (RU2); sarcoma translocation breakpoints; serine 2 (TMPRSS2) ETS fusion gene; sialyl Lewis adhesion molecule (sLe); SLAMF4; SLAMF6; Slea (CA19.9 or sialyl Lewis antigen); sperm protein 17 (SPA17); squamous cell carcinoma antigen recognized by T cells 3 (SART3); stage-specific embryonic antigen-4 (SSEA-4); STEAP1; survivin; synovial sarcoma X breakpoint 2 (SSX2); TCR gamma alternative reading frame protein (TARP); TCR-beta 1 chain; TCR-beta 2 chain; TCR-delta chain; TCR-gamma chain; TCR gamma-delta; telomerase; TGF beta R2; TNT antibody-recognized antigen; thyroid stimulating hormone receptor (TSHR); Timl- / HVCR1; tissue factor 1 (TF1); Tn ag; Tn antigen ((Tn Ag) or (GalNAca-Ser / Thr)); TNF receptor family member B cell maturation (BCMA); transglutaminase 5 (TGS5); transmembrane protease; TROP2; tumor endothelial marker 1 (TEM1 / CD248); tumor endothelial marker 7 related (TEM7R); tumor protein p53 (p53); tumor-associated glycoprotein 72 (TAG72); tyrosinase; tyrosinase-related protein 2 (TRP-2); urolipin 2 (UPK2); vascular endothelial growth factor receptor 2 (VEGFR2); V-myc avian myelocytomatosis viral oncogene neuroblastoma-derived homolog (MYCN); Wilms tumor protein (WT1); or X antigen family member 1A (XAGE1).

31. The viral particle of claim 30, wherein the target binding domain (T) binds to CD7.

32. The viral particle of claim 31, wherein the target binding domain (T) that binds to CD7 comprises a polypeptide comprising a sequence with at least 90% sequence identity to SEQ ID NO: 38 or SEQ ID NO: 39, at least 95% sequence identity to SEQ ID NO: 38 or SEQ ID NO: 39, at least 99% sequence identity to SEQ ID NO: 38 or SEQ ID NO: 39, or the sequence set forth in SEQ ID NO: 38 or SEQ ID NO:

39.

33. The viral particle of any one of claims 29-32, wherein the heterologous viral glycoprotein is a VSV-G polypeptide.

34. The viral particle of claim 33, wherein the VSV-G polypeptide comprises the sequence of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, or SEQ ID NO:

25.

35. A viral particle comprising a heterologous viral glycoprotein, a targeting moiety, and a nucleic acid molecule encoding a heterologous molecule of interest, wherein; the heterologous viral glycoprotein comprises an amino acid sequence with at least 90% identity to SEQ ID NO: 23 or SEQ ID NO: 25, at least 95% identity to SEQ ID NO: 23 or SEQ ID NO: 25, at least 99% identity to SEQ ID NO: 23 or SEQ ID NO: 25, or at least 100% identity to SEQ ID NO: 23 or SEQ ID NO: 25; the targeting moiety comprises an amino acid sequence with at least 90% identity to SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, at least 95% identity to SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, at least 99% identity to SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127, or at least 100% identity to SEQ ID NO: 98, SEQ ID NO: 126, or SEQ ID NO: 127; and wherein the nucleic acid molecule encoding a heterologous molecule of interest encodes a chimeric antigen receptor and a polypeptide comprising: i) a ligand-binding extracellular domain (ECD); ii) an immune cell activating intracellular domain (ICD); and iii) a transmembrane domain linking the ECD to the ICD.

36. The viral particle of claim 35, wherein the CAR comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO: 129, at least 90% identity to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO: 129, at least 95% identity to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO: 129, at least 99% identity to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO: 129, or at least 100% identity to SEQ ID NO: 99, SEQ ID NO: 128, or SEQ ID NO:

129.

37. The viral particle of claim 35 or 36, wherein the polypeptide encoded by the nucleic acid molecule comprises the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124, or a variant of any of the foregoing.

38. The viral particle of any one of claims 35 to 37, wherein the chimeric antigen receptor and the polypeptide are encoded by separate nucleic acid molecules.

39. The viral particle of any one of claims 35 to 37, wherein the chimeric antigen receptor and the polypeptide are encoded by the same nucleic acid molecule.

40. The viral particle of claim 39, wherein the chimeric antigen receptor and the polypeptide are separated by a sequence encoding a cleavable peptide, such as a 2A peptide.

41. A method of infecting, transfecting, or transducing a cell, the method comprising contacting the cell with a particle of any one of claims 1 to 23 or a viral particle of any one of claims 29 to 40.

42. A method of delivering one or more heterologous molecules of interest to a cell, the method comprising contacting the cell with a particle of any one of claims 1 to 23 or a viral particle of any one of claims 29 to 40, wherein the viral particle comprises a nucleic acid molecule encoding the heterologous molecule of interest.

43. The method of claim 41 or 42, wherein the method further comprises administering to the cell or to the subject one or more compounds that bind to the ECD of the polypeptide encoded by the one or more nucleic acid molecules, such as but not limited to growth hormone, G-CSF, Granocyte, Filgrastim (Neupogen, Zarzio, Nivestim, Accofil), long-acting (pegylated) Filgrastim (Pegfilgrastim, Neulasta, Pelmeg, Ziextenco), or Lenograstim (Lonquex).

44. A method of treating a disease or a condition in a subject, the method comprising administering to the subject a particle according to any one of claims 1 to 23 or a viral particle according to any one of claims 29 to 40, wherein the viral particle comprises a nucleic acid molecule encoding a heterologous molecule of interest for treating the disease or the condition, wherein the nucleic acid molecule encoding a heterologous molecule of interest is a nucleic acid molecule according to any one of claims 24 to 28.

45. A method of treating a disease or a condition in a subject, the method comprising: i) administering to the subject a particle according to any one of claims 1 to 23 or a viral particle according to any one of claims 29 to 40, and ii) administering to the subject an effective amount of one or more compounds capable of binding to the ECD of human GCSFR, thereby treating the disease or the condition, wherein the viral particle comprises one or more nucleic acid molecules encoding one or more heterologous molecules of interest for treating the disease or the condition, wherein the one or more nucleic acid molecules encoding one or more heterologous molecules of interest are one or more nucleic acid molecules according to any one of claims 24 to 28.

46. A method of treating a disease or a condition in a subject, the method comprising: i) administering to the subject a particle according to any one of claims 1 to 23 or a viral particle according to any one of claims 29 to 40, and ii) administering to the subject an effective amount of one or more compounds capable of binding to the ECD of human GHR, thereby treating the disease or the condition, wherein the viral particle comprises one or more nucleic acid molecules encoding one or more heterologous molecules of interest for treating the disease or the condition, wherein the one or more nucleic acid molecules encoding one or more heterologous molecules of interest are one or more nucleic acid molecules according to any one of claims 24 to 28.

47. A method of treating a disease or a condition in a subject, the method comprising: i) administering to the subject a viral particle according to any one of claims 29 to 40, and ii) administering to the subject an effective amount of one or more compounds capable of binding to the ECD of human GHR, thereby treating the disease or the condition, wherein the viral particle comprises a nucleic acid molecule encoding a heterologous molecule of interest for treating the disease or the condition, wherein the nucleic acid molecule or plurality of nucleic acid molecules encoding a heterologous molecule of interest encodes a chimeric antigen receptor and a polypeptide, the polypeptide comprising: i) a ligand-binding extracellular domain (ECD); ii) an immune cell activating intracellular domain (ICD); and iii) a transmembrane domain linking the ECD to the ICD.

48. The method of claim 47, wherein the CAR comprises an amino acid sequence having at least 85% identity to SEQ ID NO: 99, at least 90% identity to SEQ ID NO: 99, at least 95% identity to SEQ ID NO: 99, at least 99% identity to SEQ ID NO: 99, or at least 100% identity to SEQ ID NO:

99.

49. The method of claim 47 or 48, wherein the polypeptide encoded by the nucleic acid molecule comprises an amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 117, SEQ ID NO: 119, SEQ ID NO: 120, or SEQ ID NO: 124, or a variant of any of the foregoing.

50. A method of delivering one or more heterologous molecules to a target cell, the method comprising contacting the cell with a particle according to any one of claims 1 to 23 or a viral vector or particle according to any one of claims 29 to 40, wherein the particle comprises one or more nucleic acid molecules encoding the one or more heterologous molecules.

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