Hematopoietic cell targeting conjugates and related methods

Abstract

Provided herein are, inter alia, conjugates comprising a targeting agent (e.g., a hematopoietic cell (e.g., erythroid precursor cell) targeting agent) that comprises a protein (e.g., an antibody) that specifically binds to the transferrin receptor (TFR) (e.g., human TFR (hTFR) (e.g., hTFR1)); operably connected to (b) at least one oligonucleotide that modulates the expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by a target cell; and methods of manufacturing and pharmaceutical compositions comprising the same. Further provided herein are methods of utilizing the conjugates, including, e.g., methods of treating hemoglobinopathies (e.g., sickle cell disease (SCD) or a thalassemia (e.g., α-thalassemia, β-thalassemia, δ-thalassemia, or γ-thalassemia).

Description

Abstract This article provides, in particular, conjugates comprising a target agent (e.g., a hematopoietic cell (e.g., erythroid progenitor cell) target agent) comprising a protein (e.g., an antibody) that specifically binds to a transferrin receptor (TFR) (e.g., human TFR (hTFR) (e.g., hTFR1)); operably linked to (b) at least one oligonucleotide that regulates the expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed on the target cell; and methods for manufacturing these conjugates and pharmaceutical compositions comprising these conjugates. This article further provides methods for utilizing these conjugates, including, for example, methods for treating hemoglobinopathies (e.g., sickle cell disease (SCD)) or thalassemias (e.g., α-thalassemia, β-thalassemia, δ-thalassemia, or γ-thalassemia)).

Claims

CLAIMSWhat is claimed is:

1. A conjugate comprising:(a) a hematopoietic cell targeting agent that comprises a protein (e.g., an antibody) that specifically binds to the transferrin receptor (TFR) (e.g., human TFR (hTFR) (e.g., hTFRl)); operably connected to(b) at least one oligonucleotide that modulates (e.g., inhibits) the expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by the hematopoietic cell.

2. The conjugate of claim 1, wherein upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the conjugate is internalized into the hematopoietic cell.

3. The conjugate of claim 1 or 2, wherein the conjugate exhibits one or more of the following properties: (a) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the conjugate does not induce death of the target cell; (b) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the hematopoietic cell remains viable; (c) upon internalization into a hematopoietic cell, the conjugate does not induce death of the hematopoietic cell; and / or (d) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the conjugate does not induce degradation of TFR (e.g., hTFR (e.g., TFR1)).

4. The conjugate of any one of claims 1-3, wherein the conjugate exhibits one or more of the following properties: (a) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the conjugate is internalized into the hematopoietic cell; (b) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the conjugate does not induce death of the target cell; (c) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the hematopoietic cell remains viable; (d) upon internalization into a hematopoietic cell, the conjugate does not induce death of the hematopoietic cell; and / or (e) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the conjugate does not induce degradation of TFR (e.g., hTFR (e.g., TFR1)).

5. The conjugate of any one of claims 1-4, wherein the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) exhibits one or more of the followingproperties: (a) upon binding to TFR (e.g., hTFR (e.g., TFR 1)) expressed on the surface of a hematopoietic cell, the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) or conjugate is internalized into the hematopoietic cell; (b) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) or conjugate does not induce death of the target cell; (c) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the hematopoietic cell remains viable; (d) upon internalization into a hematopoietic cell, the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) or conjugate does not induce death of the hematopoietic cell; and / or (e) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of a hematopoietic cell, the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) or conjugate does not induce degradation of TFR (e.g., hTFR (e.g., TFR1)).

6. A conjugate comprising:(a) an erythroid precursor cell targeting agent that comprises a protein (e.g., an antibody) that specifically binds to the transl'crrin receptor (TFR) (e.g., human TFR (hTFR) (e.g., hTFRl)); operably connected to(b) at least one oligonucleotide that modulates (e.g., inhibits) the expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by the erythroid precursor cell.

7. The conjugate of claim 6, wherein upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the conjugate is internalized into the erythroid precursor cell.

8. The conjugate of claim 6 or 7, wherein the conjugate exhibits one or more of the following properties: (a) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the conjugate does not induce death of the target cell; (b) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the erythroid precursor cell remains viable; (c) upon internalization into an erythroid precursor cell, the conjugate does not induce death of the erythroid precursor cell; and / or (d) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the conjugate does not induce degradation of TFR (e.g., hTFR (e.g., TFR1)).

9. The conjugate of any one of claims 6-9, wherein the conjugate exhibits one or more of the following properties: (a) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the conjugate is internalized into the erythroid precursor cell; (b) upon binding to TFR e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the conjugate does not induce death of the target cell; (c) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the erythroid precursor cell remains viable; (d) upon internalization into an erythroid precursor cell, the conjugate does not induce death of the erythroid precursor cell; and / or (e) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the conjugate does not induce degradation of TFR (e.g., hTFR (e.g., TFR1)).

10. The conjugate of any one of claims 6-10, wherein the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR e.g., hTFRl)) exhibits one or more of the following properties: (a) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) or conjugate is internalized into the erythroid precursor cell; (b) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) or conjugate does not induce death of the target cell; (c) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the erythroid precursor cell remains viable; (d) upon internalization into an erythroid precursor cell, the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) or conjugate does not induce death of the erythroid precursor cell; and / or (e) upon binding to TFR (e.g., hTFR (e.g., TFR1)) expressed on the surface of an erythroid precursor cell, the protein (e.g., an antibody) that specifically binds to the TFR (e.g., hTFR (e.g., hTFRl)) or conjugate does not induce degradation of TFR (e.g., hTFR (e.g., TFR1)).

11. The conjugate of any one of the preceding claims, wherein the protein that specifically binds TFR (e.g., hTFR (e.g., TFR1)) is an anti-TFR (e.g., hTFR (e.g., TFR1)) antibody.

12. The conjugate of claim 11, wherein the antibody comprises or consists of a full-length antibody, a Fab, a Fab', a F(ab')2, a Fab-Fc, a scFv, a scFv-Fc, a (scFv)2-Fc, an Fv, a single domain antibody (sdAb) (e.g., a VHH), a sdAb-Fc (e.g., a VHH-Fc), a (sdAb)2 (e.g., a (VHH)2), or a (sdAb)2-Fc (e.g., a (VHH)2-Fc).

13. The conjugate of any one of claims 11 -12, wherein the antibody is an IgG (e.g., a human IgG (hlgG)) antibody.

14. The conjugate of any one of claims 11-13, wherein the antibody is a hlgGl, h!gG2, h!gG3, or h!gG4 antibody (e.g., a hlgGl or h!gG4 antibody).

15. The conjugate of any one of claims 11-14, wherein the antibody comprises an immunoglobulin (Ig) (e.g., a human Ig (hlg)) Fc region.

16. The conjugate of any one of claims 11-15, wherein the antibody comprises or consists of a full-length antibody, a Fab-Fc, a scFv-Fc, a (scFv)2-Fc, a sdAb-Fc (e.g., a VHH-Fc), or a (sdAb)2-Fc (e.g., a (VHH)2-Fc).

17. The conjugate of any one of claims 15-16, wherein the Ig (e.g., hlg) Fc region comprises at least a portion of a hinge region, a CH2 region, and a CH3 region.

18. The conjugate of any one of claims 15-17, wherein the Ig (e.g., hlg) Fc region comprises a hinge region, a CH2 region, and a CH3 region.

19. The conjugate of any one of claims 15-18, wherein the Ig is a hlg.

20. The conjugate of claim 19, wherein the hlg is a human IgG (hlgG).

21. The conjugate of claim 20, wherein the hlgG is hlgGl or hIgG4.

22. The conjugate of any one of claims 15-21, wherein the Ig (e.g., hlg) Fc region comprises one or more amino acid substitutions relative to a reference Ig (e.g., hlg) Fc region that reduces or abolishes one or more of the following effector functions relative to the reference hlg Fc region: antibody dependent cell mediated cytotoxicity (ADCC), complement dependent cytotoxicity (CDC), and / or affinity to one or more human Fc receptor (e.g., an Fey receptor (e.g., FcyRI, FcyRIIa, FcyRIIc, FcyRIIIa, and / or FcyRIIIb (e.g., FcyRI, Fcylla, and / or Fcyllla))).

23. The conjugate of any one of claims 15-22, wherein the Ig (e.g., hlg) Fc region does not substantially mediate ADCC, does not substantially mediate CDC, and / or does not bind to one or more human Fc receptor (e.g., an Fey receptor (e.g., FcyRI, FcyRIIa, FcyRIIc, FcyRIIIa, and / or FcyRIIIb (e.g., FcyRI, Fcylla, and / or Fcyllla))).

24. The conjugate of any one of claims 15-23, wherein the Ig is hlgGl and the amino acid sequence of the Fc region comprises an amino acid substitution at amino acid position L234, and / or an amino acid substitution at amino acid position L235, numbering according to the EU index of Kabat.

25. The conjugate of any one of claims 15-24, wherein the Ig is hlgGl and the amino acid sequence of the Fc region comprises an alanine at amino acid position L234 and / or an alanine at amino acid position L235, numbering according to the EU index of Kabat.

26. The conjugate of any one of claims 15-25, wherein the Ig is hlgGl and the amino acid sequence of the Fc region comprises an alanine at amino acid position L234, an alanine at amino acid position L235, and / or a glycine, an alanine, or a serine at position P329 numbering according to the EU index of Kabat.

27. The conjugate of any one of claims 15-26, wherein the Ig is hlgGl and the amino acid sequence of the Fc region comprises an alanine at amino acid position L234, a serine at amino acid position L235, and / or a glycine, an alanine, or a serine at position P329 numbering according to the EU index of Kabat.

28. The conjugate of any one of claims 15-27, wherein the Ig is hlgGl and the amino acid sequence of the Fc region comprises an alanine at amino acid position N297, numbering according to the EU index of Kabat.

29. The conjugate of any one of claims 15-23, wherein the Ig is hIgG4 and the amino acid sequence of the Fc region comprises an amino acid substitution at amino acid position S228, an amino acid substitution at amino acid position F234, and / or an amino acid substitution at amino acid position L235, numbering according to the EU index of Kabat.

30. The conjugate of any one of claims 1-23 or 29, wherein the Ig is hIgG4 and the amino acid sequence of the Fc region comprises a proline at amino acid position S228, an alanine at amino acid position F234, and / or an alanine at amino acid position L235, numbering according to EU index of Kabat.

31. The conjugate of any one of claims 15-23 or 29-30, wherein the Ig is hIgG4 and the amino acid sequence of the Fc region comprises an alanine at amino acid position N297, numbering according to the EU index of Kabat.

32. The conjugate of any one of claims 11-31, wherein the antibody comprises a first Fc region and a second Fc region that associate via at least one covalent (e.g., disulfide) bond.

33. The conjugate of any one of claims 11-32, wherein the antibody the amino acid sequence of the first Fc region and / or the amino acid sequence of the second Fc region comprise one or more amino acid substitution that promotes the association (e.g., heterodimerization) of the first and second Fc regions.

34. The conjugate of any one of claims 11 -33, wherein the amino acid sequence of the first Fc region comprises an amino acid substitution at amino acid positions T366, L368, and Y407, numbering according to the EU index of Kabat.

35. The conjugate of any one of claims 11-34, wherein the amino acid sequence of the first Fc comprises a serine at amino acid position T366, an alanine at amino acid position L368, and a valine at amino acid position Y407, numbering according to the EU index of Kabat.

36. The conjugate of any one of claims 11-35, wherein the amino acid sequence of the first Fc region comprises an amino acid substitution at amino acid position Y349, numbering according to the EU index of Kabat.

37. The conjugate of any one of claims 11-36, wherein the amino acid sequence of the first Fc region comprises a cysteine at amino acid position Y349, numbering according to the EU index of Kabat.

38. The conjugate of any one of claims 11-37, wherein the amino acid sequence of the second Fc region comprises an amino acid substitution at amino acid position T366, numbering according to the EU index of Kabat.

39. The conjugate of any one of claims 11-38, wherein the amino acid sequence of the second Fc region comprises a tryptophan at amino acid position T366, numbering according to the EU index of Kabat.

40. The conjugate of any one of claims 11-39, wherein the amino acid sequence of the second Fc region of the antibody comprises an amino acid substitution at amino acid position S354, numbering according to the EU index of Kabat.

41. The conjugate of any one of claims 11-40, wherein the amino acid sequence of the second Fc region of the antibody comprises a cysteine at amino acid position S354, numbering according to the EU index of Kabat.

42. The conjugate of any one of claims 11-41, wherein the antibody does not (or does not substantially) block binding of TF (e.g., hTF) to the TFR (e.g., hTFRl).

43. The conjugate of any one of the preceding claims, wherein the protein that specifically binds TFR (e.g., hTFR (e.g., hTFRl)) is a TFR ligand (or a functional fragment or functional variant thereof).

44. The conjugate of claim 43, wherein the TFR ligand comprises transferrin (TF) (e.g., human transferrin (hTF)) (or a functional fragment or functional variant thereof).

45. The conjugate of any one of the preceding claims, wherein the oligonucleotide enhances the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein.

46. The conjugate of any one of the preceding claims, wherein the oligonucleotide inhibits the expression and / or activity of the target gene, nucleic acid e.g., mRNA), and / or protein.

47. The conjugate of any one of the preceding claims, wherein the oligonucleotide modulates (e.g., enhances or inhibits) the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein through binding to a target nucleic acid molecule encoded by the target gene, nucleic acid (e.g., mRNA), and / or protein (e.g., target mRNA molecule (e.g., a portion of a target mRNA molecule)).

48. The conjugate of any one of the preceding claims, wherein the target nucleic acid molecule is a target mRNA molecule (e.g., a portion of a target mRNA molecule).

49. The conjugate of any one of the preceding claims, wherein the oligonucleotide mediates one or more of the following degradation of the target nucleic acid molecule (e.g., mRNA), disabling of the target nucleic acid molecule (e.g., mRNA), modification of the target nucleic acid molecule (e.g., mRNA), alteration in the splicing of the target nucleic acid molecule (e.g., mRNA), alteration (e.g., a decrease) in the stability of the target nucleic acid molecule (e.g., mRNA), or a block in the translation of the target nucleic acid molecule (e.g., mRNA), or any combination of the foregoing.

50. The conjugate of any one of the preceding claims, wherein the oligonucleotide comprises or consists of an antisense oligonucleotide (ASO), small interfering RNA (siRNA), a short hairpin RNA (shRNA), or a microRNA (miRNA).

51. The conjugate of any one of the preceding claims, wherein the oligonucleotide comprises or consists of an antisense strand comprising a region of complementarity to a target sequence (e.g., an mRNA sequence encoded by the target gene, nucleic acid (e.g., mRNA), and / or protein).

52. The conjugate of any one of the preceding claims, wherein the oligonucleotide is single stranded or double stranded.

53. The conjugate of any one of the preceding claims, wherein the oligonucleotide is a DNA, RNA, or RNA and RNA hybrid molecule.

54. The conjugate of any one of the preceding claims, wherein the oligonucleotide comprises a sense strand and an antisense strand forming a double stranded region.

55. The conjugate of any one of the preceding claims, wherein the sense strand and the antisense strand arc part of a single nucleic acid molecule (e.g., wherein a hairpin loop is between the sense strand and the antisense strand of the single nucleic acid molecule.

56. The conjugate of any one of the preceding claims, wherein the sense strand and the antisense strand are separate nucleic acid molecules (i.e., connected only through the double stranded region).

57. The conjugate of any one of the preceding claims, wherein the double stranded region is from about 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-20, 19-21, 23-30, 23-29, 23-28, 23-27, 23-26, 23-25, 23-24, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21- 23, or 21-22 nucleotide pairs in length.

58. The conjugate of any one of the preceding claims, wherein the oligonucleotide comprises at least one modified nucleotide.

59. The conjugate of any one of the preceding claims, wherein at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the nucleotides of the oligonucleotide are modified.

60. The conjugate of any one of the preceding claims, wherein substantially all (or all) of the nucleotides in the oligonucleotide are modified.

61. The conjugate of any one of the preceding claims, wherein at least one of the modified nucleotides comprises a modified sugar (e.g., ribose moiety).

62. The conjugate of any one of the preceding claims, wherein at least one of the modified nucleotides comprises a modified nucleobase.

63. The conjugate of any one of the preceding claims, wherein the oligonucleotide comprises at least one modified internucleoside linkage (e.g., at least one phosphorothioate intemucleoside linkage).

64. The conjugate of any one of the preceding claims, wherein the at least one modified nucleotide is a 2’ modified nucleotide (e.g., a 2'-fluoro (2'-F), 2'-O-methyl (2'-0-Me), 2'-O- methoxyethyl (2'-M0E), 2'-O- aminopropyl (2'-O-AP), 2'-O-dimethylaminoethyl (2'-0-DMA0E), 2'-O-dimethylaminopropyl (2'-0-DMAP), 2'-O-dimethylaminoethyloxyethyl (2'-O-DMAEOE), 2'-O-N-methylacetamido (2'-0-NMA), locked nucleic acid (LNA), ethylene -bridged nucleic acid (ENA), and (S)- constrained ethyl-bridged nucleic acid (cEt) (e.g., a 2' modified nucleotide is 2'- O-methyl or 2'-fluoro (2'-F))).

65. The conjugate of any one of the preceding claims, wherein the protein encoded by the target gene, nucleic acid (e.g., mRNA), and / or protein is associated with one or more hemoglobinopathy .

66. The conjugate of any one of the preceding claims, wherein inhibition of or a reduction in expression and / or activity of a protein encoded by the target gene, nucleic acid (e.g., mRNA), and / or protein is associated with an increase in the level of fetal hemoglobin, the induction of expression of fetal hemoglobin, and / or an increase in the ratio of fetal hemoglobin to adult hemoglobin.

67. The conjugate of any one of the preceding claims, wherein expression and / or activity of a protein encoded by the target gene, nucleic acid (e.g., mRNA), and / or protein is associated with a repression of fetal hemoglobin, a decrease in the level of fetal hemoglobin, an increase in the level of adult hemoglobin, and / or an increase in ratio of adult hemoglobin to fetal hemoglobin.

68. The conjugate of any one of the preceding claims, wherein the target gene, nucleic acid (e.g., mRNA), and / or protein is a transcription factor.

69. The conjugate of any one of the preceding claims, wherein the target gene, nucleic acid (e.g., mRNA), and / or protein is highly expressed in erythroid precursor cells (relative to other non-erythroid precursor cell types).

70. The conjugate of any one of the preceding claims, wherein the target gene is B cell lymphoma leukemia 11A (BCL11A) (e.g., human BCL11A (e.g., hBCLUA)), Zinc Finger and BTB Domain Containing 7A (ZBTB7A) (e.g., hZBTB7A), KLF Transcription Factor 1 (KLF1) (e.g., hKLFl), FA Complementation Group A (FANCA) (e.g., human FANCA), Dyskerin Pseudouridine Synthase 1 (DKC1) (e.g., human DKC1), Regulator of Telomere Elongation Helicase 1 (RTEL1) (e.g., human RTEL1), Telomerase Reverse Transcriptase (TERT) (e.g., human TERT), Telomerase RNA Component (TERC) (e.g., human TERC), TERFI Interacting Nuclear Factor 2 (TINF2) (e.g., human TINF2), Ribosomal Protein S19 (RPS19) (e.g., human RPS19), Ribosomal Protein Li l (RPL11) (e.g., human RPL11), Ribosomal Protein S26 (RPS26) (e.g., human RPS26), Ribosomal Protein S10 (RPS10) (e.g., human RPS10), Ribosomal Protein L35A (RPL35A) (e.g., human RPL35A), Ribosomal Protein S24 (RPS24) (e.g., human RPS24), Ribosomal Protein S17 (RPS17) (e.g., human RPS17), SBDS Ribosome Maturation Factor (SBDS) (e.g., human SBDS), Signal Recognition Particle 54 (SRP54) (e.g., human SRP54), E74 Like ETS Transcription Factor 1 (ELF1) (e.g., human ELF1), Elastase Neutrophil Expressed(ELA2) (e.g., human ELA2), HCLS1 Associated Protein X-l (HAX1 ) (e.g., human HAX1), Glucosc-6-Phosphatasc Catalytic Subunit 3 (G6PC3) (e.g., human G6PC3), Growth Factor Independent 1 Transcriptional Repressor (GFI1) e.g., human GFI1), WASP Actin Nucleation Promoting Factor (WAS) (e.g., human WAS), Colony Stimulating Factor 3 Receptor (CSF3R) (e.g., human CSF3R), MPE Proto-Oncogene Thrombopoietin Receptor (MPL) (e.g., human MPL), GATA Binding Protein 2 (GATA2) (e.g., human GATA2), Sterile Alpha Motif Domain Containing 9 (SAMD9) (e.g., human SAMD9), Sterile Alpha Motif Domain Containing 9 Eike (SAMD9L) (e.g., human SAMD9L), or MDS1 And EVI1 Complex Locus (MECOM) (e.g., human MECOM).

71. The conjugate of any one of the preceding claims, wherein the target gene is BCL11A (e.g., hBCLUA), ZBTB7A (e.g., hZBTB7A), or KLF1 (e.g., hKLFl).

72. The conjugate of any one of the preceding claims, wherein (a) the protein that specifically binds TFR is non-covalently conjugated to (b) the at least one oligonucleotide.

73. The conjugate of any one of the preceding claims, wherein (a) the protein that specifically binds TFR is covalently conjugated to (b) the at least one oligonucleotide.

74. The conjugate of any one of the preceding claims, wherein (a) the protein that specifically binds TFR is directly conjugated to (b) the at least one oligonucleotide.

75. The conjugate of any one of the preceding claims, wherein (a) the protein that specifically binds TFR is indirectly conjugated to (b) the at least one oligonucleotide through (c) a linker.

76. The conjugate of any one of the preceding claims, wherein the linker is cleavable or non- cleavable.

77. The conjugate of any one of the preceding claims, wherein the wherein (b) comprises at least 2, 3, 4, 5, 6, or more oligonucleotides.

78. The conjugate of any one of the preceding claims, wherein each of the at least 2, 3, 4, 5, 6, or more oligonucleotides are individually conjugated to (a) the protein (e.g., antibody) that specifically binds TFR (e.g., as described herein).

79. A cell comprising the conjugate of any one of claims 1-78.

80. The cell of claim 79, wherein the cell is in vitro, ex vivo, or in vivo.

81. A pharmaceutical composition comprising the conjugate of any one of claims 1-78 and a pharmaceutically acceptable excipient.

82. A kit comprising the conjugate of any one of claims 1 -78 or the pharmaceutical composition of any one of claims 81.

83. A method of delivering a conjugate or pharmaceutical composition to a cell, the method comprising introducing into a cell the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby deliver the conjugate or pharmaceutical composition into the cell.

84. The method of claim 83, wherein the cell is in vitro, ex vivo, or in vivo.

85. The method of claim 83 or 84, wherein the cell is a subject (e.g., a human subject).

86. A method of delivering a conjugate, cell, or pharmaceutical composition to a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78, the cell of any one of claims 79-80, or the pharmaceutical composition of claim 81, to thereby deliver the conjugate, cell, or pharmaceutical composition to the subject.

87. A method of modulating (e.g., inhibiting or enhancing) expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by a hematopoietic cell in a cell, the method comprising introducing into the cell the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby modulate (e.g., inhibit or enhance) the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein.

88. A method of modulating (e.g., inhibiting or enhancing) expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by an erythroid precursor cell in a cell, the method comprising introducing into the cell the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby modulate (e.g., inhibit or enhance) the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein.

89. The method of any one of claims 87-88, wherein the cell is in vitro, ex vivo, or in vivo.

90. The method of any one of claims 87-89, wherein the cell is a subject (e.g., a human subject).

91. A method of modulating (e.g., inhibiting or enhancing) expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by a hematopoietic cell in a cell in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby modulate (e.g., inhibit or enhance) the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein in the subject.

92. A method of modulating (e.g., inhibiting or enhancing) expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by an erythroid precursor cell in a cell in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby modulate (e.g., inhibit or enhance) the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein in the subject.

93. A method of reducing and / or inhibiting expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by a hematopoietic cell in a cell, the method comprising introducing into the cell the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby reduce or inhibit the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein.

94. A method of reducing and / or inhibiting expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by an erythroid precursor cell in a cell, the method comprising introducing into the cell the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby reduce or inhibit the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein.

95. The method of any one of claims 93-94, wherein the cell is in vitro, ex vivo, or in vivo.

96. The method of any one of claims 93-95, wherein the cell is a subject (e.g., a human subject).

97. The method of any one of claims 93-96, wherein the target is BCL11 A (e.g., hBCLl 1 A), ZBTB7A (e.g., hZBTB7A), or KLF1 (e.g., hKLFl).

98. A method of reducing and / or inhibiting expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by a hematopoietic cell in a cell in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby reduce or inhibit the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein in the subject.

99. A method of reducing and / or inhibiting expression and / or activity of a target gene, nucleic acid (e.g., mRNA), and / or protein expressed by an erythroid precursor cell in a cell in a subject, the method comprising administering to the subject the conjugate of any one of claims 1- 78 or the pharmaceutical composition of claim 81, to thereby reduce or inhibit the expression and / or activity of the target gene, nucleic acid (e.g., mRNA), and / or protein in the subject.

100. A method of inducing expression of fetal hemoglobin in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby induce expression of fetal hemoglobin the subject.

101. A method of increasing the level of fetal hemoglobin in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby increase the level of fetal hemoglobin the subject.

102. A method of increasing the ratio of fetal hemoglobin to adult hemoglobin in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby increase the ratio of fetal hemoglobin to adult hemoglobin in the subject.

103. A method of treating, ameliorating, or preventing an inherited blood disorder in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby treat, ameliorate, or prevent the inherited blood disorder in the subject.

104. The method of claim 103, wherein the inherited blood disorder is a hemoglobinopathy or an inherited bone marrow failure syndrome.

105. A method of treating, ameliorating, or preventing a hemoglobinopathy in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby treat, ameliorate, or prevent the hemoglobinopathy in the subject.

106. The method of claim 105, wherein the hemoglobinopathy is sickle cell disease, sickle cell trait, hemoglobin C disease, hemoglobin C trait, hemoglobin S / C disease, hemoglobin D disease, hemoglobin E disease, a thalassemia (e.g., a-thalassemia, P-thalassemia, 8-thalassemia, or y- thalassemia), a condition associated with hemoglobin with increased oxygen affinity, a condition associated with hemoglobin with decreased oxygen affinity, unstable hemoglobin disease, methemoglobinemia, or any combination thereof.

107. The method of claim 105 or 106, wherein the hemoglobinopathy is sickle cell disease or a thalassemia (e.g., a-thalassemia, P-thalassemia, 8-thalassemia, or y-thalassemia).

108. The method of any one of claims 105-107, wherein the subject is a human.

109. The method of any one of claims 105-108, wherein the subject is suspected of having or has been diagnosed with sickle cell disease, sickle cell trait, hemoglobin C disease, hemoglobinC trait, hemoglobin S / C disease, hemoglobin D disease, hemoglobin E disease, a thalassemia (e.g., a-thalasscmia, P-thalasscmia, 8-thalasscmia, or ' / -thalassemia), a condition associated with hemoglobin with increased oxygen affinity, a condition associated with hemoglobin with decreased oxygen affinity, unstable hemoglobin disease, methemoglobinemia, or any combination thereof.

110. The method of any one of claims 105-109, wherein the subject is suspected of having or has been diagnosed with sickle cell disease or a thalassemia (e.g., a- thalassemia, P-thalassemia, 6-thalassemia, or ' / -thalassemia).

111. A method of treating, ameliorating, or preventing an inherited bone marrow failure syndrome in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-78 or the pharmaceutical composition of claim 81, to thereby treat, ameliorate, or prevent the inherited bone marrow failure syndrome in the subject.

112. The method of claim 111, wherein the inherited bone marrow failure syndrome is amegakaryocytic thrombocytopenia (Amega), diamond blackfan anemia (DBA), dyskeratosis congenita (DC), Fanconi anemia (FA), Pearson syndrome, severe congenital neutropenia (SCN), Schwachman diamond syndrome (SDS), GATA2 deficiency, cyclic neutropenia, Dubowitz syndrome, Kostmann syndrome, refractory cytopenia, thrombocytopenia absent radii (TAR) a SAMD9 / SAMD9E disorder, or a MECOM-associated syndrome.

113. The conjugate of any one of claims 1-78, the cell of any one of claims 79-80, or the pharmaceutical composition of claim 81 for use in the treatment of a disease in a subject in need thereof.

114. The conjugate of any one of claims 1-78, the cell of any one of claims 79-80, or the pharmaceutical composition of claim 81 for use as a medicament.

115. Use of the conjugate of any one of claims 1-78, the cell of any one of claims 79-80, or the pharmaceutical composition of claim 81 for the manufacture of a medicament for the treatment of a disease in a subject in need thereof.

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