Methods for enhancing lifespan and/or treating cellular proliferative disorders by transplantation
The invention found that first, the feasibility of transfer of tumor resistance and other healthy longevity characters through transplantation of bone marrow mononuclear cells (BMMNC) or hematopoietic stem cells (HSC)/hematopoietic stem and progenitor cells (HSPC) consisting of genetically engineered EKLF gene encoding the hematopoietic transcription factor EKLF. Secondly, the present invention demonstrates expression of EKLF in the long-term hematopoietic stem cells (LT-HSC), and thus EKLF as a target of regulation of hematopoiesis.
1 . A method of increasing longevity and/or inhibiting or reducing tumor occurrence or tumor metastasis of a subject, comprising:
(a) genetically engineering cord blood stem cells (CBSCs) or bone marrow mononuclear cells (BMMNCs) to possess one or more modified Erythroid Kruppel-like factor (Eklf) genes encoding a modified human EKLF polypeptide comprising an amino acid substitution at the sumoylation site that corresponds to lysine at position 54 of the wild type human EKLF, or a modified mouse EKLF polypeptide comprising an amino acid substitution at the sumoylation site that corresponds to lysine at position 74 of the wild type mouse EKLF; and
(b) transplanting the genetically engineered CBSCs or BMMNCs to the subject.
2 . The method of claim 1 , wherein the CBSCs or BMMNCs are cord blood hematopoietic cells, peripheral hematopoietic cells, hematopoietic stem cells (HSCs), hematopoietic stem and progenitor cells (HSPCs), multipotent progenitors (MPPs), common myeloid progenitors (CMPs), granulocyte/macrophage progenitors (GMPs), myeloid/erythroid progenitors (MEPs), lymphoid cells, or myeloid cells.
3 . The method of claim 1 , wherein the modified mouse EKLF polypeptide comprises a substitution of the lysine (K) residue corresponding to position 74 of the wild type mouse EKLF with an arginine (R) or with another amino acid that confers tumor resistance and healthy longevity.
4 . The method of claim 1 , wherein the modified human EKLF polypeptide comprises a substitution of the lysine (K) residue corresponding to position 54 of the wild type human EKLF with an arginine (R) or with another amino acid that confers tumor resistance and healthy longevity.
5 . The method of claim 1 , wherein the modified mouse EKLF polypeptide comprises an amino acid substitution at position 68 of the full length wild-type mouse EKLF polypeptide.
6 . The method of claim 1 , wherein the CBSCs or BMMNCs are transduced to express the modified EKLF polypeptide via use of a viral vector encoding the modified EKLF polypeptide.
7 . The method of claim 6 , wherein the viral vector is derived from a herpes virus, a retrovirus, a vaccinia virus, an attenuated vaccinia virus, a canary pox virus, an adenovirus, or an adeno-associated virus.
8 . The method of claim 1 , wherein the CBSCs or BMMNCs are transduced to express the modified human EKLF polypeptide or the modified mouse EKLF polypeptide via use of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR associated proteins (Cas) system.
9 . The method of claim 1 , wherein the expression of the modified human EKLF polypeptide or the modified mouse EKLF polypeptide leads to enhanced lifespan, anti-metastasis, and/or anti-tumorigenesis.
10 . The method of claim 2 , wherein the modified human EKLF polypeptide or the modified mouse EKLF polypeptide is expressed at a relatively high level in long-term (LT)-HSCs and depletion of EKLF leads to population changes of different types of hematopoietic cells or hematopoietic progenitors.
11 . The method of claim 2 , wherein the modified human EKLF polypeptide or the modified mouse EKLF polypeptide negatively regulates the expression of colony-stimulating factor 2 receptor subunit Csf2rb in LT-HSCs and the hematopoietic cells or the hematopoietic progenitors.
12 . The method of claim 1 , wherein the tumor is liver cancer, colon cancer, breast cancer, prostate cancer, hepatocellular carcinoma, melanoma, lung cancer, glioblastoma, brain tumor, hematopoietic malignancies, retinoblastoma, renal cell carcinoma, head and neck cancer, cervical cancer, pancreatic cancer, esophageal cancer, or squamous cell carcinoma.