Compositions and methods for selective elimination and replacement of hematopoietic stem cells
Disclosed are methods of eliminating at least on target cell in a subject, comprising administering to the subject an effective amount of a composition comprising a plurality of immune cells, wherein each immune cell of the plurality expresses one or more chimeric ligand receptor(s) (CLR(s)) that each specifically bind to a target ligand on the at least one target cell, wherein specifically binding of the one or more CLR(s) to the target activates the immune cell, and wherein the activated immune cell induces death of the target cell. Exemplary target cells include, but are not limited to, hematopoietic stem cells (HSCs).
1. A method of eliminating at least one target hematopoietic stem cell (HSC) in a subject, comprising administering to the subject an effective amount of a composition comprising a plurality of T-cells that express at least one chimeric ligand receptor (CLR) that specifically binds to c-KIT,
wherein the specific binding of the at least one CLR to c-KIT activates the T-cell,
wherein the activated T-cell induces death of the at least one target HSC, and
wherein the at least one CLR comprises:
(a) an ectodomain comprising
i) a signal peptide comprising a human CD8α signal peptide;
ii) a scFv comprising the amino acid sequence of SEQ ID NO: 73; and
iii) a hinge domain comprising a human CD8α hinge domain;
(b) a transmembrane domain comprising a human CD8α transmembrane domain;
(c) a co-stimulatory domain comprising a human 4-1BB costimulatory domain; and
(d) a endodomain comprising a human CD3ζ endodomain.
2. The method of claim 1 , further comprising eliminating the plurality of T-cells following the death of the at least one target cell.
3. The method of claim 2 , further comprising administering to the subject an effective amount of a composition comprising a plurality of therapeutic hematopoietic stem cells (HSCs) after eliminating the plurality of T-cells.
4. The method of claim 1 , wherein the plurality of T-cells in the composition comprises resting CAR-T-cells, activated CAR-T cells, or a combination thereof.
5. The method of claim 1 , wherein the composition comprises a plurality of allogeneic T-cells.
6. The method of claim 1 , wherein the composition comprises a plurality of autologous T-cells.
7. The method of claim 1 , further comprising administering to the subject an effective amount of a preconditioning composition to enhance engraftment of the composition comprising the plurality of T-cells.
8. The method of claim 7 , wherein the preconditioning composition suppresses the immune system of the subject.
9. The method of claim 1 , wherein the subject has or is at risk of developing an immune system disease or disorder, is immunocompromised, has an autoimmune disease or disorder, has an inflammatory disease or disorder, or any combination thereof.
10. The method of claim 1 , wherein the subject has a genetic or epigenetic marker for a disease or disorder that manifests in a blood cell, an immune cell circulating in the blood, a bone marrow cell or a precursor cell thereof.
11. The method of claim 1 , wherein the subject has an anemia, a clotting disorder or a hemorrhagic condition.
12. The method of claim 1 , wherein the subject has cancer.
13. The method of claim 12 , wherein the cancer is a lymphoma, a leukemia, a myeloma or a malignant immunoproliferative disease.
14. The method of claim 1 , wherein i) the signal peptide comprises the amino acid sequence of SEQ ID NO: 31.
15. The method of claim 1 , wherein iii) a hinge domain comprises the amino acid sequence of SEQ ID NO: 40.
16. The method of claim 1 , wherein (b) the transmembrane domain comprises the amino acid of SEQ ID NO: 33.
17. The method of claim 1 , wherein (c) the first costimulatory domain comprises the amino acid of SEQ ID NO: 38.
18. The method of claim 1 , wherein (d) the endodomain comprises the amino acid sequence of SEQ ID NO: 36.
19. The method of claim 1 , wherein
(a) the ectodomain comprises
i) a signal peptide comprising the amino acid sequence of SEQ ID NO: 31;
ii) an scFv comprising the amino acid sequence of SEQ ID NO: 73;
iii) a hinge domain comprising the amino acid sequence of SEQ ID NO: 40;
(b) the transmembrane domain comprises the amino acid of SEQ ID NO: 33;
(c) the first costimulatory domain comprises the amino acid of SEQ ID NO: 38; and
(d) the endodomain comprises the amino acid sequence of SEQ ID NO: 36.