COMPOSITIONS AND METHODS FOR REPROGRAMMING HEMATOPOIETIC STEM CELL LINEAGES

• US 20160032317A1
• Filed: 03/14/2014
• Published: 02/04/2016
• Est. Priority Date: 03/14/2013
• Status: Abandoned Application

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Medical Center Corporation
• Inventors: Rossi, Derrick, Riddell, Jonah, Gazit, Roi
• Application Number:
  US14/774,785
• Publication Number:
  US 20160032317A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: C07K 2319/50 containing protease site
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2506/00 Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
• CPC Class Codes: C12N 2506/11 from blood or immune system cells
• CPC Class Codes: C12N 2506/1307 from adult fibroblasts
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2730/00041 Use of virus, viral particle or viral elements as a vector
• CPC Class Codes: C12N 2740/16043 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2830/003 tet inducible
• CPC Class Codes: C12N 2840/20 translation of more than one cistron
• CPC Class Codes: C12N 5/0647 Haematopoietic stem cells; Uncommitted or multipotent progenitors

Abstract

Provided herein are compositions, methods, and kits for hematopoietic stem cell induction or for reprogramming cells to the multipotent state of hematopoietic stem cells. In some embodiments, the compositions comprise at least one HSC inducing factor. Such compositions, methods and kits can be used for inducing hematopoietic stem cells in vitro, ex vivo, or in vivo, as described herein, and these induced hematopoietic stem cells can be used in regenerative medicine applications and therapies.

Claims

1. A hematopoietic stem cell (HSC) inducing composition comprising one or more expression vector encoding four or more HSC inducing factors selected from:

CDKN1C, DNMT3B, EGR1, ETV6, EVI1, GATA2, GFI1B, GLIS2, HLF, HMGA2, HOXA5, HOXA9, HOXB3, HOXB4, HOXB5, IGF2BP2, IKZF2, KLF12, KLF4, KLF9, LMO2, MEIS1, MSI2, MYCN, NAP1L3, NDN, NFIX, NKX2-3, NR3C2, PBX1, PRDM16, PRDM5, RARB, RBBP6, RBPMS, RUNX1, RUNX1T1, SMAD6, TAL1, TCF15, VDR, ZFP37, ZFP467, ZFP521, ZFP532, ZFP612, and ZPF467.

48. The HSC inducing composition of claim 1, wherein the four or more HSC inducing factors are HLF, RUNX1T1, PBX1, LMO2, PRDM5, ZFP37, MYCN, MSI2, NKX2-3, MEIS1, and RBPMS.

49. The HSC inducing composition of claim 1, wherein the four or more HSC inducing factors are HLF, RUNX1T1, ZFP37, PBX1, LMO2, and PRDM5.

50. The HSC inducing compositions of claim 1, wherein the expression vector is a retroviral vector.

51. The HSC inducing compositions of claim 1, wherein the expression vector is a lentiviral vector.

52. The HSC inducing composition of claim 51, wherein the lentiviral vector is an inducible lentiviral vector.

68. A kit for making induced hematopoietic stem cells (iHSCs) comprising the HSC inducing compositions of claim 1.

2-47. -47. (canceled)

53. A method for preparing an induced hematopoietic stem cell (iHSC) from a somatic cell comprising:

a. transducing the somatic cell with one or more vectors comprising a nucleic acid sequence encoding HLF, a nucleic acid sequence encoding RUNX1T1;

a nucleic acid sequence encoding ZFP37;

a nucleic acid sequence encoding PBX1;

a nucleic acid sequence encoding LMO2; and

a nucleic acid sequence encoding PRDM5, wherein each said nucleic acid sequence is operably linked to a promoter; and

b. culturing the transduced somatic cell in a cell media that supports growth of hematopoietic stem cells, thereby preparing an iHSC.

54. The method of claim 53, wherein the transducing of step (a) further comprises one or more vectors comprising one or more of:

a nucleic acid sequence encoding PRDM16 a nucleic acid sequence encoding ZFP467; and

a nucleic acid sequence encoding VDR wherein each said nucleic acid sequence is operably linked to a promoter.

55. The method of claim 53 further comprising transducing the somatic cell with one or more vectors comprising a nucleic acid sequence encoding ZFP37;

a nucleic acid sequence encoding MYCN;

a nucleic acid sequence encoding MSI2;

a nucleic acid sequence encoding NKX2-3;

a nucleic acid sequence encoding MEIS1; and

a nucleic acid sequence encoding RBPMS;

wherein each said nucleic acid sequence is operably linked to a promoter.

56. The method of claim 53, wherein the expression vector is a retroviral vector.

57. The method of claim 53, wherein the expression vector is a lentiviral vector.

58. The method of claim 57, wherein the lentiviral vector is an inducible lentiviral vector.

59. The method of claim 53, wherein the somatic cell is a fibroblast cell.

60. The method of claim 53, wherein the somatic cell is a hematopoietic lineage cell.

61. The method of claim 60, wherein the hematopoietic lineage cell is selected from promyelocytes, neutrophils, eosinophils, basophils, reticulocytes, erythrocytes, mast cells, osteoclasts, megakaryoblasts, platelet producing megakaryocytes, platelets, monocytes, macrophages, dendritic cells, lymphocytes, NK cells, NKT cells, innate lymphocytes, multipotent hematopoietic progenitor cells, oligopotent hematopoietic progenitor cells, and lineage restricted hematopoietic progenitors.

62. The method of claim 60, wherein the hematopoietic lineage cell is selected from a multi-potent progenitor cell (MPP), common myeloid progenitor cell (CMP), granulocyte-monocyte progenitor cells (GMP), common lymphoid progenitor cell (CLP), and pre-megakaryocyte-erythrocyte progenitor cell.

63. The method of claim 60, wherein the hematopoietic lineage cell is selected from a megakaryocyte-erythrocyte progenitor cell (MEP), a ProB cell, a PreB cell, a PreProB cell, a ProT cell, a double-negative T cell, a pro-NK cell, a pro-dendritic cell (pro-DC), pre-granulocyte/macrophage cell, a granulocyte/macrophage progenitor (GMP) cell, and a pro-mast cell (ProMC).

66. The method of claim 63, wherein the expression vector is a retroviral vector.

67. The method of claim 63, wherein the expression vector is a lentiviral vector.

64. A method of promoting transdifferentiation of a ProPreB cell to the myeloid lineage comprising:

a. transducing a ProPreB cell with one or more vectors comprising a nucleic acid sequence encoding ZFP467, a nucleic acid sequence encoding PBX1;

a nucleic acid sequence encoding HOXB4; and

a nucleic acid sequence encoding MSI2;

wherein each said nucleic acid sequence is operably linked to a promoter; and

b. culturing the transduced ProPreB cell in a cell media that supports growth of myeloid lineage cells, thereby transdifferentiating the ProPreB cell to the myeloid lineage.

65. The method of claim 64, wherein the transducing of step (a) further comprises one or more vectors comprising one or more of:

a nucleic acid sequence encoding HLF, a nucleic acid sequence encoding LMO2;

a nucleic acid sequence encoding PRDM16; and

a nucleic acid sequence encoding ZFP37.