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US-20260125645-A1 - METHODS AND PHARMACOLOGICAL AGENTS FOR INCREASING EXPANSION, ENGRAFTMENT OR IMMUNE OUTPUT OF HUMAN HEMATOPOIETIC STEM CELLS IN TRANSPLANTATION AND DISEASE SETTINGS

US20260125645A1US 20260125645 A1US20260125645 A1US 20260125645A1US-20260125645-A1

Abstract

The present application relates to methods for expanding hematopoietic stem cells (HSC) and/or hematopoietic progenitor cells (HPC) ex vivo and/or in vivo. The methods comprise culturing the HSCs and/or HPCs in the presence of a selective 5-Hydroxytryptamine Receptor 1F (HTR1F) agonist and/or of a Thyroid Hormone Receptor Beta (THRB) agonist, and/or in an air-liquid interface (ALI) culture system. The expanded HSCs and/or HPCs, which maintain engraftment and immune reconstitution properties, may be administered to subjects suffering from various diseases including cancer, immunodeficiencies, myelodysplastic syndrome (MDS) or anemia. The present application also relates to the use of HTR1F and/or THRB agonists for stimulating HSC and/or HPC expansion, and for increasing the number of immune cells or for reconstituting the immune system, in vivo.

Inventors

  • Élie HADDAD
  • Panojot BIFSHA

Assignees

  • VALORISATION HSJ, LIMITED PARTNERSHIP

Dates

Publication Date
20260507
Application Date
20231003

Claims (20)

  1. 1 . A method for expanding hematopoietic stem cells (HSCs) and/or hematopoietic progenitor cells (HPCs) ex vivo comprising culturing said hematopoietic stem cells and/or hematopoietic progenitor cells under conditions suitable for HSC and/or HPC expansion (a) in the presence of a selective 5-Hydroxytryptamine Receptor 1F (HTR1F) agonist and/or of a Thyroid Hormone Receptor Beta (THRB) agonist; and/or (b) in an air-liquid interface (ALI) culture system.
  2. 2 . The method of claim 1 , wherein the HSCs and/or HPCs are obtained from donor bone marrow, umbilical donor cord blood, or donor peripheral blood.
  3. 3 . The method of claim 1 , wherein the HSCs and/or HPCs are from a subject suffering from a disease or condition.
  4. 4 . The method of claim 3 , wherein the disease is cancer, an immunodeficiency, myelodysplastic syndrome (MDS) or anemia.
  5. 5 . The method of claim 4 , wherein the cancer is a hematological cancer.
  6. 6 . The method of claim 1 , further comprising genetically modifying the HSCs and/or HPCs to generate genetically-modified HSCs and/or HPCs.
  7. 7 . The method of claim 1 , wherein said method comprises culturing the HSCs and/or HPCs in an ALI culture system.
  8. 8 . The method of claim 1 , wherein said conditions suitable for HSC and/or HPC expansion comprises culturing the HSCs and/or HPCs in the presence of cytokines and/or growth factors.
  9. 9 . The method of claim 8 , wherein the cytokines and/or growth factors comprises SCF, TPO, FLT3L and/or IL-6.
  10. 10 . (canceled)
  11. 11 . The method of claim 1 , wherein the selective HTR1F agonist is N-[(3R)-3-(Dimethylamino)-2,3,4,9-tetrahydro-1H-carbazol-6-yl]-4-fluorobenzamide hydrochloride (LY 344864), 4-fluoro-N-(3-(1-methyl-4-piperidinyl)-1H-indol-5-yl)benzamide (LY 334370), 2,4,6-Trifluoro-N-[6-[(1-methyl-4-piperidinyl)carbonyl]-2-pyridinyl]benzamide (LY 573144, Lasmiditan), or 5-N-butyryloxy-N,N-dimethyltryptamine.
  12. 12 . The method of claim 1 , wherein the THRB agonist is triiodothyronine (T3).
  13. 13 - 20 . (canceled)
  14. 21 . A method for (i)expanding hematopoietic stem cells (HSCs) and/or hematopoietic progenitor cells (HPCs) and/or (ii) increasing the number of immune cells or for reconstituting the immune system in a subject in need thereof comprising administering to the subject an effective amount of a selective 5-Hydroxytryptamine Receptor 1F (HTR1F) agonist and/or of a Thyroid Hormone Receptor Beta (THRB) agonist.
  15. 22 . The method of claim 21 , wherein the selective HTR1F agonist is N-[(3R)-3-(Dimethylamino)-2,3,4,9-tetrahydro-1H-carbazol-6-yl]-4-fluorobenzamide hydrochloride (LY 344864), 4-fluoro-N-(3-(1-methyl-4-piperidinyl)-1H-indol-5-yl)benzamide (LY 334370), 2,4,6-Trifluoro-N-[6-[(1-methyl-4-piperidinyl)carbonyl]-2-pyridinyl]benzamide (LY 573144, Lasmiditan), or 5-N-butyryloxy-N,N-dimethyltryptamine.
  16. 23 . The method of claim 21 , wherein the THRB agonist is triiodothyronine (T3).
  17. 24 . The method of claim 21 , wherein the subject is a stem-cell transplant recipient.
  18. 25 . The method of claim 21 , wherein the subject suffers from cancer, an immunodeficiency, myelodysplastic syndrome (MDS) or anemia.
  19. 26 . The method of claim 25 , wherein the cancer is a hematological cancer.
  20. 27 . The method of claim 24 , wherein the stem-cell transplant comprises hematopoietic stem cells (HSCs) and/or hematopoietic progenitor cells (HPCs) obtained by an ex vivo comprising culturing said HSCs and/or HPCs under conditions suitable for HSC and/or HPC expansion (a) in the presence of a selective 5-Hydroxytryptamine Receptor 1F (HTR1F) agonist and/or of a Thyroid Hormone Receptor Beta (THRB) agonist; and/or (b) in an air-liquid interface (ALI) culture system.

Description

CROSS REFERENCE TO RELATED APPLICATIONS The present application claims the benefit of U.S. provisional patent application No. 63/412,944 filed on Oct. 4, 2022, which is incorporated herein by reference in its entirety. TECHNICAL FIELD The present invention generally relates to the field of hematology, and more particularly to stem cell transplantation and treatment of blood or immunological disorders. BACKGROUND ART Hematopoietic stem cells (HSCs) promote the lifelong production of all mature blood cell lineages through their unique capabilities of durable self-renewal and multilineage differentiation. HSCs are present in donor-derived bone marrow (BM), cord blood (CB), and mobilized peripheral blood stem-cell products for allogeneic stem-cell transplantation (SCT) in patients with hematological malignancies and monogenic diseases. Patient-derived peripheral blood stem-cell products are extensively used for autologous SCT, which supports hematopoietic rescue after high-dose chemotherapy for various types of hematological malignancies, solid tumors, and autoimmune diseases. However, in many diseases affecting blood cell lineages, the number of HSCs that can be obtained from a patient is very limited in number. Attrition of autologous HSCs during processing for gene therapy is also a problem. In addition, it has not been possible to maintain or expand human HSCs ex vivo. Under in vitro cell culture conditions, the number of HCS typically decline because HSCs either die or terminally differentiate, losing their stem cell properties. In many situations, the clinical outcome would be much better if one could expand or amplify the number, in the graft, of self-renewing and multipotent HSC capable of reconstituting immune system and maintaining hematopoiesis long-term.1 Problems associated with current ex vivo stem cell expansion methods include loss of short-term repopulating activity of cultured cells and expansion of aberrant long-term repopulating cells with poor contribution to peripheral immune reconstitution. Studies have shown that aryl hydrocarbon receptor antagonist2, such as StemRegenin1 (SR1), notch ligand agonists3, and UM171/7294 can all promote HSC expansion and increase repopulating capacities, particularly from cord blood. Attempting to replicate state-of-the-art methodologies that use either SR1 or UM171/729, for the expansion of primitive HSC, led to the observation that these protocols commonly failed to maintain short-term repopulating activity in humanized mice experiments5. Furthermore, even though these culture methods reported a significant increase of long-term repopulating activity, they lacked robustness and were rarely outperforming the non-cultured HSC as to their ability to populate the bone-marrow and peripheral blood of immunodeficient mouse hosts. In clinical transplantation, the presence of sufficient short-term repopulating activity (stemming from progenitors and primitive HSC) is of paramount importance for the survival of the patient. Indeed, this short-term repopulating activity is responsible for the rapid increase of polymorphonuclear and platelets, which is crucial after transplantation. Also, the continuous lifetime production of blood cells is required and can only be maintained by primitive HSC6. There is thus a need in the fields of HSCT and gene therapy for the development of strategies that would increase the numbers of engraftable primitive HSC without skewing the engraftment characteristics of the cells of origin7-10 to enhance regeneration of the blood system, notably in the treatment of hematological malignancies, solid tumors and autoimmune diseases. The present description refers to a number of documents, the content of which is herein incorporated by reference in their entirety. SUMMARY The present disclosure provides the following items 1 to 42: 1. A method for expanding hematopoietic stem cells (HSCs) and/or hematopoietic progenitor cells (HPCs) ex vivo comprising culturing said hematopoietic stem cells and/or hematopoietic progenitor cells in the presence of a selective 5-Hydroxytryptamine Receptor 1F (HTR1F) agonist and/or of a Thyroid Hormone Receptor Beta (THRB) agonist under conditions suitable for HSC and/or HPC expansion.2. The method of item 1, wherein the HSCs and/or HPCs are obtained from donor bone marrow, umbilical donor cord blood, or donor peripheral blood.3. The method of item 1 or 2, wherein the HSCs and/or HPCs are from a subject suffering from a disease or condition.4. The method of item 3, wherein the disease is cancer, an immunodeficiency, myelodysplastic syndrome (MDS) or anemia.5. The method of item 4, wherein the cancer is a hematological cancer.6. The method of any one of items 1 to 5, further comprising genetically modifying the HSCs and/or HPCs to generate genetically-modified HSCs and/or HPCs.7. The method of any one of items 1 to 6, wherein said conditions suitable for HSC and/or HPC expansion comprises culturing the HSCs and/or H