Composition and use method therefor

Abstract

An aging reprogramming composition, a use method therefor, a kit containing the composition, and the use of the composition in tissue repair, tissue regeneration, organ regeneration, aging reversion, or any combination thereof. The present invention relates to SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1 and c-MYC, which are used as inducing factors to reprogram cells, tissues or organs to restore a youthful state. Provided is a composition of an activation inducing factor in the form of an engineered nucleic acid, an engineered cell, an engineered protein, a chemical agent, or any combination thereof, wherein the composition is used for treating diseases, preventing diseases, modulating tissue repair, modulating tissue regeneration, or any combination thereof.

Description

Compositions and methods of use Technical Field

[0001] The present invention relates to a highly efficient method for inducing aging reprogramming of somatic cells, and also provides a reprogramming composition for rejuvenating cells, tissues or organs of a subject. Background Art

[0002] All organisms experience aging, which is accompanied by a decline in cellular and tissue function. Aging is an extremely complex process characterized by a time-dependent decline in function at the molecular, cellular, tissue, and organismal levels. Cellular senescence is not the same as cell death; while senescent cells retain metabolic activity for a period of time, they also exhibit several distinct changes: typical epigenetic alterations, genomic instability, telomere loss, loss of protein homeostasis, mitochondrial dysfunction, stem cell depletion, altered intercellular communication, chronic inflammation, and more than a dozen other hallmarks.

[0003] Cell reprogramming technology involves modulating and altering cell fate without altering the cell's genetic sequence through various methods, such as the introduction of exogenous genes or the treatment of small molecules. This includes improving or even reversing cellular aging. However, existing combinations of reprogramming factors that can achieve anti-aging or cell vitality restoration are relatively limited. Therefore, there is an urgent need for more effective combinations of reprogramming factors that can restore the vitality of a subject's cells, tissues, or organs, or combat aging.

[0004] Summary of the Invention

[0005] The present invention relates to a composition for rejuvenating cells, tissues, or organs in a subject, and a method for rejuvenating cells, tissues, or organs in a subject using the composition. The composition and method are capable of rejuvenating or restoring the function of cells, tissues, or organs in a subject. The composition and method can be used for any disease or injury requiring the repair or replacement of dysfunctional tissue.

[0006] In a first aspect, the present invention provides a composition for rejuvenating cells, tissues or organs of a subject, the composition comprising: any one factor selected from SOX1, SOX2, SOX3, RCOR2, GMNN and GLIS1; any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4 and PAX1; factor Klf4 or Klf2; and / or, factor c-MYC.

[0007] In some embodiments, the composition comprises: any one factor selected from SOX1, SOX2, SOX3, RCOR2 and GMNN; any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4 and PAX1; factor Klf4 or Klf2; and / or, factor c-MYC.

[0008] In some embodiments, the composition comprises: any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, NKX3.1, FOXA2, NR5A2, OCT4, and SOX7; a KLF2 or KLF4 factor; any one factor selected from SOX1, SOX2, GLIS1, or SOX3; and / or, a factor c-MYC.

[0009] In some embodiments, the composition comprises GATA6, SOX3, and Klf4.

[0010] In some embodiments, the composition comprises OCT4, SOX3, and Klf4.

[0011] In some embodiments, the composition comprises PAX1, SOX3, and Klf4.

[0012] In some embodiments, the composition comprises GATA3, SOX3, and Klf4.

[0013] In some embodiments, the composition comprises C\EBPa, SOX3, and Klf4.

[0014] In some embodiments, the composition comprises OCT4, SOX1, and Klf4.

[0015] In some embodiments, the composition comprises GATA3, SOX1, and Klf4.

[0016] In some embodiments, the composition comprises GATA6, SOX1, and Klf4.

[0017] In some embodiments, the composition comprises SOX17, SOX1, and Klf4.

[0018] In some embodiments, the composition comprises NR5A2, SOX2, and Klf4.

[0019] In some embodiments, the composition comprises GATA1, SOX2, and Klf4.

[0020] In some embodiments, the composition comprises GATA2, SOX2, and Klf4.

[0021] In some embodiments, the composition comprises GATA3, SOX2, and Klf4.

[0022] In some embodiments, the composition comprises GATA4, SOX2, and Klf4.

[0023] In some embodiments, the composition comprises GATA6, SOX2, and Klf4.

[0024] In some embodiments, the composition comprises FOXA2, SOX2, and Klf4.

[0025] In some embodiments, the composition comprises SOX7, SOX2, and Klf4.

[0026] In some embodiments, the composition comprises PAX1, SOX2, and Klf4.

[0027] In some embodiments, the composition comprises SOX17, SOX2, and Klf4.

[0028] In some embodiments, the composition comprises C\EBPa, SOX2, and Klf4.

[0029] In some embodiments, the composition comprises NKX3.1, SOX2, and Klf4.

[0030] In some embodiments, the composition comprises GATA6, GMNN, and Klf4.

[0031] In some embodiments, the composition comprises OCT4, RCOR2, and Klf4.

[0032] In some embodiments, the composition comprises OCT4, SOX2, and Klf2.

[0033] In some embodiments, the composition comprises OCT4, GMNN, and Klf2.

[0034] In some embodiments, the composition comprises OCT4, RCOR2, and Klf2.

[0035] In some embodiments, the composition comprises FOXA2, SOX2, and Klf2.

[0036] In some embodiments, the composition comprises GATA1, SOX1, and KLF4.

[0037] In some embodiments, the composition comprises GATA2, SOX1, and KLF4.

[0038] In some embodiments, the composition comprises GATA4, SOX1, and KLF4.

[0039] In some embodiments, the composition comprises FOXA2, SOX1, and KLF4.

[0040] In some embodiments, the composition comprises NKX3.1, SOX1, and KLF4.

[0041] In some embodiments, the composition comprises GATA1, SOX3, and KLF4.

[0042] In some embodiments, the composition comprises GATA2, SOX3, and KLF4.

[0043] In some embodiments, the composition comprises GATA4, SOX3, and KLF4.

[0044] In some embodiments, the composition comprises FOXA2, SOX3, and KLF4.

[0045] In some embodiments, the composition comprises NKX3.1, SOX3, and KLF4.

[0046] In some embodiments, the composition comprises SOX7, SOX3, and KLF4.

[0047] In some embodiments, the composition comprises NR5A2, SOX3, and KLF4.

[0048] In some embodiments, the composition comprises GATA1, SOX1, and KLF2.

[0049] In some embodiments, the composition comprises GATA2, SOX1, and KLF2.

[0050] In some embodiments, the composition comprises GATA3, SOX1, and KLF2.

[0051] In some embodiments, the composition comprises GATA4, SOX1, and KLF2.

[0052] In some embodiments, the composition comprises GATA6, SOX1, and KLF2.

[0053] In some embodiments, the composition comprises NKX3.1, SOX1, and KLF2.

[0054] In some embodiments, the composition comprises OCT4, SOX1, and KLF2.

[0055] In some embodiments, the composition comprises GATA3, SOX2, and KLF2.

[0056] In some embodiments, the composition comprises GATA4, SOX2, and KLF2.

[0057] In some embodiments, the composition comprises GATA6, SOX2, and KLF2.

[0058] In some embodiments, the composition comprises GATA1, SOX3, and KLF2.

[0059] In some embodiments, the composition comprises GATA2, SOX3, and KLF2.

[0060] In some embodiments, the composition comprises GATA6, SOX3, and KLF2.

[0061] In some embodiments, the composition comprises OCT4, SOX3, and KLF2.

[0062] In some embodiments, the composition comprises KLF4, OCT4, and GLIS1.

[0063] In some embodiments, the composition comprises KLF2, OCT4, and GLIS1.

[0064] In some embodiments, the composition comprises GATA6, SOX3, Klf4, and c-MYC.

[0065] In some embodiments, the composition comprises GATA3, SOX3, Klf4, and c-MYC.

[0066] In some embodiments, the composition comprises OCT4, SOX3, Klf4, and c-MYC.

[0067] In some embodiments, the composition comprises PAX1, SOX3, Klf4, and c-MYC.

[0068] In some embodiments, the composition comprises C\EBPa, SOX3, Klf4, and c-MYC.

[0069] In some embodiments, the composition comprises OCT4, SOX1, Klf4, and c-MYC.

[0070] In some embodiments, the composition comprises GATA3, SOX1, Klf4, and c-MYC.

[0071] In some embodiments, the composition comprises GATA6, SOX1, Klf4, and c-MYC.

[0072] In some embodiments, the composition comprises SOX17, SOX1, Klf4, and c-MYC.

[0073] In some embodiments, the composition comprises NR5A2, SOX2, Klf4, and c-MYC.

[0074] In some embodiments, the composition comprises GATA1, SOX2, Klf4, and c-MYC.

[0075] In some embodiments, the composition comprises GATA2, SOX2, Klf4, and c-MYC.

[0076] In some embodiments, the composition comprises GATA3, SOX2, Klf4, and c-MYC.

[0077] In some embodiments, the composition comprises GATA4, SOX2, Klf4, and c-MYC.

[0078] In some embodiments, the composition comprises GATA6, SOX2, Klf4, and c-MYC.

[0079] In some embodiments, the composition comprises FOXA2, SOX2, Klf4, and c-MYC.

[0080] In some embodiments, the composition comprises SOX7, SOX2, Klf4, and c-MYC.

[0081] In some embodiments, the composition comprises PAX1, SOX2, Klf4, and c-MYC.

[0082] In some embodiments, the composition comprises SOX17, SOX2, Klf4, and c-MYC.

[0083] In some embodiments, the composition comprises C\EBPa, SOX2, Klf4, and c-MYC.

[0084] In some embodiments, the composition comprises NKX3.1, SOX2, Klf4, and c-MYC.

[0085] In some embodiments, the composition comprises GATA6, GMNN, Klf4, and c-MYC.

[0086] In some embodiments, the composition comprises OCT4, RCOR2, Klf4, and c-MYC.

[0087] In some embodiments, the composition comprises OCT4, SOX2, Klf2, and c-MYC.

[0088] In some embodiments, the composition comprises OCT4, GMNN, Klf2, and c-MYC.

[0089] In some embodiments, the composition comprises OCT4, RCOR2, Klf2, and c-MYC.

[0090] In some embodiments, the composition comprises FOXA2, SOX2, Klf2, and c-MYC.

[0091] In some embodiments, the composition comprises GATA1, SOX1, KLF4, and c-MYC.

[0092] In some embodiments, the composition comprises GATA2, SOX1, KLF4, and c-MYC.

[0093] In some embodiments, the composition comprises GATA4, SOX1, KLF4, and c-MYC.

[0094] In some embodiments, the composition comprises FOXA2, SOX1, KLF4, and c-MYC.

[0095] In some embodiments, the composition comprises NKX3.1, SOX1, KLF4, and c-MYC.

[0096] In some embodiments, the composition comprises GATA1, SOX3, KLF4, and c-MYC.

[0097] In some embodiments, the composition comprises GATA2, SOX3, KLF4, and c-MYC.

[0098] In some embodiments, the composition comprises GATA4, SOX3, KLF4, and c-MYC.

[0099] In some embodiments, the composition comprises FOXA2, SOX3, KLF4, and c-MYC.

[0100] In some embodiments, the composition comprises NKX3.1, SOX3, KLF4, and c-MYC.

[0101] In some embodiments, the composition comprises SOX7, SOX3, KLF4, and c-MYC.

[0102] In some embodiments, the composition comprises NR5A2, SOX3, KLF4, and c-MYC.

[0103] In some embodiments, the composition comprises GATA1, SOX1, KLF2, and c-MYC.

[0104] In some embodiments, the composition comprises GATA2, SOX1, KLF2, and c-MYC.

[0105] In some embodiments, the composition comprises GATA3, SOX1, KLF2, and c-MYC.

[0106] In some embodiments, the composition comprises GATA4, SOX1, KLF2, and c-MYC.

[0107] In some embodiments, the composition comprises GATA6, SOX1, KLF2, and c-MYC.

[0108] In some embodiments, the composition comprises NKX3.1, SOX1, KLF2, and c-MYC.

[0109] In some embodiments, the composition comprises OCT4, SOX1, KLF2, and c-MYC.

[0110] In some embodiments, the composition comprises GATA3, SOX2, KLF2, and c-MYC.

[0111] In some embodiments, the composition comprises GATA4, SOX2, KLF2, and c-MYC.

[0112] In some embodiments, the composition comprises GATA6, SOX2, KLF2, and c-MYC.

[0113] In some embodiments, the composition comprises GATA1, SOX3, KLF2, and c-MYC.

[0114] In some embodiments, the composition comprises GATA2, SOX3, KLF2, and c-MYC.

[0115] In some embodiments, the composition comprises GATA6, SOX3, KLF2, and c-MYC.

[0116] In some embodiments, the composition comprises OCT4, SOX3, KLF2, and c-MYC.

[0117] In some embodiments, the composition comprises KLF4, OCT4, GLIS1, and c-MYC.

[0118] In some embodiments, the composition comprises KLF2, OCT4, GLIS1, and c-MYC.

[0119] In some embodiments, the factor is a protein, a nucleic acid encoding the same, or a mixture thereof.

[0120] For example, SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, and c-MYC are proteins, fusion proteins bound to a membrane-permeable domain, and encode nucleic acids, such as in the form of DNA or RNA.

[0121] In some embodiments, the factor is provided in the form of an expression vector, which can be a plasmid vector, an episomal vector, a transposon, a virus-derived vector, such as an adenovirus vector, an adeno-associated virus vector (AAV), a lentivirus vector, a Sendai virus vector, a cytomegalovirus vector or a vaccinia virus vector.

[0122] In some embodiments, the factors are present on one or more expression vectors.

[0123] In some embodiments, the factors are present on the same expression vector.

[0124] In some embodiments, the factor is in the form of mRNA.

[0125] In some embodiments, the factors are tandemly present on the same mRNA molecule, or each of the factors is present on a different mRNA molecule, and the factors are in the form of a complex of multiple mRNA molecules.

[0126] In some embodiments, the factor is a synthetic mRNA encoding a wild-type form, a mutant that retains its activity, or a genetically engineered form of a gene selected from the group consisting of SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, and c-MYC.

[0127] In some embodiments, the composition further comprises a pharmaceutically acceptable carrier or adjuvant.

[0128] In some embodiments, the subject is a mammal, including but not limited to humans, mice, rats, cows, sheep, horses, dogs, cats, pigs, or monkeys.

[0129] In some embodiments, the subject is a chicken, duck, goose, or bird.

[0130] In some embodiments, the subject cell, tissue or organ is from, but is not limited to, the eye, ear, nose, mouth, including gums and tooth roots; bone, lung, breast, pancreas, stomach, esophagus; muscle, including myocardium; liver, blood vessels; skin, including hair; heart, brain, nervous tissue, kidney, testicle, prostate, penis, cloaca, fin, ovary or intestine.

[0131] In some embodiments, the composition for rejuvenating cells, tissues, or organs of a subject is capable of restoring epigenetic information to the cells, tissues, or organs of the subject.

[0132] In some embodiments, the composition for rejuvenating cells, tissues, or organs in a subject is capable of restoring epigenetic information lost in the cells, tissues, or organs of the subject due to aging, injury, or disease.

[0133] In some embodiments, the expression of promoter regulatory factors can be used to reduce and reverse epigenetic marks associated with aging, increase epigenetic marks associated with cell rejuvenation, reduce the expression of aging-associated proteins, increase the expression of proteins associated with cell rejuvenation, restore the balance between euchromatin and heterochromatin, prevent the loss of cell identity, restore cell identity, reverse aging-associated DNA methylation changes, and thus rejuvenate cells.

[0134] In some embodiments, the SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC is a human protein, or a non-human protein, for example, a mammalian (e.g., mouse, rat, cow, sheep, horse, dog, cat, pig, monkey, chicken, duck, goose, bird) protein, or a protein that has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity with the amino acid sequence of the corresponding human protein or non-human protein and maintains activity.

[0135] In some embodiments, the SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC, or any combination thereof is a nucleic acid encoding a human protein, or a nucleic acid encoding a non-human protein, for example, a nucleic acid encoding a protein of a mammal (e.g., mouse, rat, cow, sheep, horse, dog, cat, pig, monkey, chicken, duck, goose, bird), or a nucleic acid that has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity to a corresponding nucleic acid encoding a human protein, or a nucleic acid sequence encoding a non-human protein and retains activity.

[0136] In some embodiments, rejuvenating a cell, tissue, or organ of a subject comprises restoring a transcriptional profile or epigenetic information lost in the at least one cell, tissue, or organ due to aging, injury, disease, or any combination thereof.

[0137] In some embodiments, rejuvenating the subject's cells, tissues, or organs comprises restoring the function of the cells, increasing the potency of the cells, enhancing the viability of the cells, or increasing the replicative capacity or lifespan of the cells, or a combination thereof.

[0138] In some embodiments, the rejuvenation of the subject's cells, tissues, or organs comprises promoting axon regeneration in the subject's neuronal cells; reversing the epigenetic age of aging in the subject's skin fibroblasts; reversing the aging of the subject's skin fibroblasts; reversing the aging of the subject's chondrocytes; reversing the aging of the subject's muscle stem cells; promoting improvement in the subject's osteoarthritis symptoms; or promoting improvement in the subject's eye disease.

[0139] In some embodiments, the subject has, is suspected of having, or is at risk of having:

[0140] Diseases, conditions or manifestations associated with impaired cell, tissue or organ vitality or aging;

[0141] The disease, condition or manifestation associated with impaired cell, tissue or organ vitality or aging is a neurodegenerative disease, cardiovascular disease, metabolic disease, musculoskeletal disease, inflammatory disease, manifestation or condition associated with the skin, eye disease, fibrotic disease, or disease associated with the reproductive system;

[0142] The neurodegenerative disease is cerebral stroke, Huntington's disease, senile dementia, Alzheimer's disease, Parkinson's disease;

[0143] The cardiovascular disease is hypertension, coronary heart disease arrhythmia, heart disease, hypotension, heart failure, angina pectoris, arteriosclerosis, myocardial infarction, myocarditis, congestive heart failure, atrioventricular block, atherosclerosis, myocardial infarction;

[0144] The metabolic disease is type I diabetes, type II diabetes, hyperglycemia, hyperinsulinemia, insulin resistance, obesity, gout;

[0145] The musculoskeletal disease is muscular dystrophy, myasthenia gravis, osteoporosis, osteoarthritis, osteochondrosis, osteochondritis, osteonecrosis, and osteopenia;

[0146] The inflammatory disease is systemic lupus erythematosus, polymyalgia rheumatica, gouty arthritis, degenerative arthritis, rheumatoid arthritis, inflammatory arthritis, Hashimoto's thyroiditis, inflammatory bowel disease, hepatitis, pneumonia, respiratory tract inflammation, encephalitis;

[0147] The skin-related aging symptoms or diseases are wrinkles, age spots, seborrheic keratosis, tinea pedis, urticaria, neurodermatitis, and pigmented nevi;

[0148] The eye disease is macular degeneration, retinal degeneration, retinal detachment, diabetic retinopathy, glaucoma, optic atrophy, floaters, cataract, non-arteritic anterior ischemic optic neuropathy, chorioretinitis, retinitis pigmentosa, optic nerve damage;

[0149] The fibrotic disease is pulmonary fibrosis, liver fibrosis, myocardial fibrosis, renal interstitial fibrosis, myelofibrosis, pleural fibrosis, intestinal wall fibrosis;

[0150] Diseases related to the reproductive system include erectile dysfunction and premature ovarian failure.

[0151] In some embodiments, the composition induces cell reprogramming, reverses aging, improves tissue function, improves organ function, promotes tissue repair, promotes tissue survival, promotes tissue regeneration, promotes tissue growth, promotes angiogenesis, reduces scar formation, alleviates external manifestations of aging including alopecia, hair thinning, hair graying, skin sagging and skin wrinkling, promotes organ regeneration, promotes organ survival, treats disease, or any combination thereof.

[0152] In some embodiments, the composition reverses aging of a cell, tissue, organ, or subject, thereby rejuvenating the cell, tissue, or organ.

[0153] In some embodiments, the composition can promote neuronal regeneration in a subject.

[0154] In some embodiments, the composition can protect neurons in a subject from damage or degeneration.

[0155] In some embodiments, the composition can reverse the epigenetic age of senescent skin fibroblasts in a subject.

[0156] In some embodiments, the composition can reverse senescence of skin fibroblasts in a subject.

[0157] In some embodiments, the composition can reverse senescence of chondrocytes in a subject.

[0158] In some embodiments, the composition can reverse aging of muscle stem cells in a subject.

[0159] In some embodiments, the composition can promote improvement of osteoarthritis symptoms in a subject.

[0160] In some embodiments, the composition can promote improvement of glaucoma symptoms in a subject.

[0161] In some embodiments, the composition can promote regeneration of the optic nerve.

[0162] In some embodiments, the composition can promote axonal regeneration in the optic nerve.

[0163] In some embodiments, the composition can inhibit apoptosis of optic nerve cells (eg, retinal ganglion cells) in optic nerve damage.

[0164] In some embodiments, the composition does not induce teratoma formation or does not induce tumor growth or tumor formation.

[0165] In some embodiments, the composition does not induce complete reprogramming.

[0166] In some embodiments, the composition does not reprogram cells into induced pluripotent stem cells (iPSCs).

[0167] In a second aspect, the present invention provides a method for rejuvenating cells, tissues or organs in a subject, the method comprising administering the composition of the first aspect to the subject.

[0168] In some embodiments, the composition comprises: any one factor selected from SOX1, SOX2, SOX3, RCOR2, GMNN and GLIS1; any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4 and PAX1; factor Klf4 or Klf2; and / or, factor c-MYC.

[0169] In some embodiments, the composition comprises: any one factor selected from SOX1, SOX2, SOX3, RCOR2 and GMNN; any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4 and PAX1; factor Klf4 or Klf2; and / or, factor c-MYC.

[0170] In some embodiments, the composition comprises: any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, NKX3.1, FOXA2, NR5A2, OCT4, and SOX7; a KLF2 or KLF4 factor; any one factor selected from SOX1, SOX2, GLIS1, or SOX3; and / or, a factor c-MYC.

[0171] In some embodiments, the composition comprises GATA6, SOX3, and Klf4.

[0172] In some embodiments, the composition comprises OCT4, SOX3, and Klf4.

[0173] In some embodiments, the composition comprises PAX1, SOX3, and Klf4.

[0174] In some embodiments, the composition comprises GATA3, SOX3, and Klf4.

[0175] In some embodiments, the composition comprises C\EBPa, SOX3, and Klf4.

[0176] In some embodiments, the composition comprises OCT4, SOX1, and Klf4.

[0177] In some embodiments, the composition comprises GATA3, SOX1, and Klf4.

[0178] In some embodiments, the composition comprises GATA6, SOX1, and Klf4.

[0179] In some embodiments, the composition comprises SOX17, SOX1, and Klf4.

[0180] In some embodiments, the composition comprises NR5A2, SOX2, and Klf4.

[0181] In some embodiments, the composition comprises GATA1, SOX2, and Klf4.

[0182] In some embodiments, the composition comprises GATA2, SOX2, and Klf4.

[0183] In some embodiments, the composition comprises GATA3, SOX2, and Klf4.

[0184] In some embodiments, the composition comprises GATA4, SOX2, and Klf4.

[0185] In some embodiments, the composition comprises GATA6, SOX2, and Klf4.

[0186] In some embodiments, the composition comprises FOXA2, SOX2, and Klf4.

[0187] In some embodiments, the composition comprises SOX7, SOX2, and Klf4.

[0188] In some embodiments, the composition comprises PAX1, SOX2, and Klf4.

[0189] In some embodiments, the composition comprises SOX17, SOX2, and Klf4.

[0190] In some embodiments, the composition comprises C\EBPa, SOX2, and Klf4.

[0191] In some embodiments, the composition comprises NKX3.1, SOX2, and Klf4.

[0192] In some embodiments, the composition comprises GATA6, GMNN, and Klf4.

[0193] In some embodiments, the composition comprises OCT4, RCOR2, and Klf4.

[0194] In some embodiments, the composition comprises OCT4, SOX2, and Klf2.

[0195] In some embodiments, the composition comprises OCT4, GMNN, and Klf2.

[0196] In some embodiments, the composition comprises OCT4, RCOR2, and Klf2.

[0197] In some embodiments, the composition comprises FOXA2, SOX2, and Klf2.

[0198] In some embodiments, the composition comprises GATA1, SOX1, and KLF4.

[0199] In some embodiments, the composition comprises GATA2, SOX1, and KLF4.

[0200] In some embodiments, the composition comprises GATA4, SOX1, and KLF4.

[0201] In some embodiments, the composition comprises FOXA2, SOX1, and KLF4.

[0202] In some embodiments, the composition comprises NKX3.1, SOX1, and KLF4.

[0203] In some embodiments, the composition comprises GATA1, SOX3, and KLF4.

[0204] In some embodiments, the composition comprises GATA2, SOX3, and KLF4.

[0205] In some embodiments, the composition comprises GATA4, SOX3, and KLF4.

[0206] In some embodiments, the composition comprises FOXA2, SOX3, and KLF4.

[0207] In some embodiments, the composition comprises NKX3.1, SOX3, and KLF4.

[0208] In some embodiments, the composition comprises SOX7, SOX3, and KLF4.

[0209] In some embodiments, the composition comprises NR5A2, SOX3, and KLF4.

[0210] In some embodiments, the composition comprises GATA1, SOX1, and KLF2.

[0211] In some embodiments, the composition comprises GATA2, SOX1, and KLF2.

[0212] In some embodiments, the composition comprises GATA3, SOX1, and KLF2.

[0213] In some embodiments, the composition comprises GATA4, SOX1, and KLF2.

[0214] In some embodiments, the composition comprises GATA6, SOX1, and KLF2.

[0215] In some embodiments, the composition comprises NKX3.1, SOX1, and KLF2.

[0216] In some embodiments, the composition comprises OCT4, SOX1, and KLF2.

[0217] In some embodiments, the composition comprises GATA3, SOX2, and KLF2.

[0218] In some embodiments, the composition comprises GATA4, SOX2, and KLF2.

[0219] In some embodiments, the composition comprises GATA6, SOX2, and KLF2.

[0220] In some embodiments, the composition comprises GATA1, SOX3, and KLF2.

[0221] In some embodiments, the composition comprises GATA2, SOX3, and KLF2.

[0222] In some embodiments, the composition comprises GATA6, SOX3, and KLF2.

[0223] In some embodiments, the composition comprises OCT4, SOX3, and KLF2.

[0224] In some embodiments, the composition comprises KLF4, OCT4, and GLIS1.

[0225] In some embodiments, the composition comprises KLF2, OCT4, and GLIS1.

[0226] In some embodiments, the composition comprises GATA6, SOX3, Klf4, and c-MYC.

[0227] In some embodiments, the composition comprises GATA3, SOX3, Klf4, and c-MYC.

[0228] In some embodiments, the composition comprises OCT4, SOX3, Klf4, and c-MYC.

[0229] In some embodiments, the composition comprises PAX1, SOX3, Klf4, and c-MYC.

[0230] In some embodiments, the composition comprises C\EBPa, SOX3, Klf4, and c-MYC.

[0231] In some embodiments, the composition comprises OCT4, SOX1, Klf4, and c-MYC.

[0232] In some embodiments, the composition comprises GATA3, SOX1, Klf4, and c-MYC.

[0233] In some embodiments, the composition comprises GATA6, SOX1, Klf4, and c-MYC.

[0234] In some embodiments, the composition comprises SOX17, SOX1, Klf4, and c-MYC.

[0235] In some embodiments, the composition comprises NR5A2, SOX2, Klf4, and c-MYC.

[0236] In some embodiments, the composition comprises GATA1, SOX2, Klf4, and c-MYC.

[0237] In some embodiments, the composition comprises GATA2, SOX2, Klf4, and c-MYC.

[0238] In some embodiments, the composition comprises GATA3, SOX2, Klf4, and c-MYC.

[0239] In some embodiments, the composition comprises GATA4, SOX2, Klf4, and c-MYC.

[0240] In some embodiments, the composition comprises GATA6, SOX2, Klf4, and c-MYC.

[0241] In some embodiments, the composition comprises FOXA2, SOX2, Klf4, and c-MYC.

[0242] In some embodiments, the composition comprises SOX7, SOX2, Klf4, and c-MYC.

[0243] In some embodiments, the composition comprises PAX1, SOX2, Klf4, and c-MYC.

[0244] In some embodiments, the composition comprises SOX17, SOX2, Klf4, and c-MYC.

[0245] In some embodiments, the composition comprises C\EBPa, SOX2, Klf4, and c-MYC.

[0246] In some embodiments, the composition comprises NKX3.1, SOX2, Klf4, and c-MYC.

[0247] In some embodiments, the composition comprises GATA6, GMNN, Klf4, and c-MYC.

[0248] In some embodiments, the composition comprises OCT4, RCOR2, Klf4, and c-MYC.

[0249] In some embodiments, the composition comprises OCT4, SOX2, Klf2, and c-MYC.

[0250] In some embodiments, the composition comprises OCT4, GMNN, Klf2, and c-MYC.

[0251] In some embodiments, the composition comprises OCT4, RCOR2, Klf2, and c-MYC.

[0252] In some embodiments, the composition comprises FOXA2, SOX2, Klf2, and c-MYC.

[0253] In some embodiments, the composition comprises GATA1, SOX1, KLF4, and c-MYC.

[0254] In some embodiments, the composition comprises GATA2, SOX1, KLF4, and c-MYC.

[0255] In some embodiments, the composition comprises GATA4, SOX1, KLF4, and c-MYC.

[0256] In some embodiments, the composition comprises FOXA2, SOX1, KLF4, and c-MYC.

[0257] In some embodiments, the composition comprises NKX3.1, SOX1, KLF4, and c-MYC.

[0258] In some embodiments, the composition comprises GATA1, SOX3, KLF4, and c-MYC.

[0259] In some embodiments, the composition comprises GATA2, SOX3, KLF4, and c-MYC.

[0260] In some embodiments, the composition comprises GATA4, SOX3, KLF4, and c-MYC.

[0261] In some embodiments, the composition comprises FOXA2, SOX3, KLF4, and c-MYC.

[0262] In some embodiments, the composition comprises NKX3.1, SOX3, KLF4, and c-MYC.

[0263] In some embodiments, the composition comprises SOX7, SOX3, KLF4, and c-MYC.

[0264] In some embodiments, the composition comprises NR5A2, SOX3, KLF4, and c-MYC.

[0265] In some embodiments, the composition comprises GATA1, SOX1, KLF2, and c-MYC.

[0266] In some embodiments, the composition comprises GATA2, SOX1, KLF2, and c-MYC.

[0267] In some embodiments, the composition comprises GATA3, SOX1, KLF2, and c-MYC.

[0268] In some embodiments, the composition comprises GATA4, SOX1, KLF2, and c-MYC.

[0269] In some embodiments, the composition comprises GATA6, SOX1, KLF2, and c-MYC.

[0270] In some embodiments, the composition comprises NKX3.1, SOX1, KLF2, and c-MYC.

[0271] In some embodiments, the composition comprises OCT4, SOX1, KLF2, and c-MYC.

[0272] In some embodiments, the composition comprises GATA3, SOX2, KLF2, and c-MYC.

[0273] In some embodiments, the composition comprises GATA4, SOX2, KLF2, and c-MYC.

[0274] In some embodiments, the composition comprises GATA6, SOX2, KLF2, and c-MYC.

[0275] In some embodiments, the composition comprises GATA1, SOX3, KLF2, and c-MYC.

[0276] In some embodiments, the composition comprises GATA2, SOX3, KLF2, and c-MYC.

[0277] In some embodiments, the composition comprises GATA6, SOX3, KLF2, and c-MYC.

[0278] In some embodiments, the composition comprises OCT4, SOX3, KLF2, and c-MYC.

[0279] In some embodiments, the composition comprises KLF4, OCT4, GLIS1, and c-MYC.

[0280] In some embodiments, the composition comprises KLF2, OCT4, GLIS1, and c-MYC.

[0281] In some embodiments, the factor is a polypeptide, a nucleic acid encoding the same, or a mixture thereof.

[0282] For example, SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, and c-MYC are proteins, and fusion proteins bound to their membrane-permeable domains, which encode nucleic acids, such as in the form of DNA or RNA.

[0283] In some embodiments, the factor is provided in the form of an expression vector, which can be a plasmid vector, an episomal vector, a transposon, a virus-derived vector, such as an adenovirus vector, an adeno-associated virus vector (AAV), a lentivirus vector, a Sendai virus vector, a cytomegalovirus vector or a vaccinia virus vector.

[0284] In some embodiments, the factors are present on one or more expression vectors.

[0285] In some embodiments, the factors are present on the same expression vector.

[0286] In some embodiments, the factor is in the form of mRNA.

[0287] In some embodiments, the factors are tandemly present on the same mRNA molecule, or each of the factors is present on a different mRNA molecule, and the factors are in the form of a complex of multiple mRNA molecules.

[0288] In some embodiments, the factor is a synthetic mRNA encoding a wild-type form, a mutant that retains its activity, or a genetically engineered form of a gene selected from the group consisting of SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, and c-MYC.

[0289] In some embodiments, the composition further comprises a pharmaceutically acceptable carrier or adjuvant.

[0290] In some embodiments, the subject is a mammal, including but not limited to humans, mice, rats, cows, sheep, horses, dogs, cats, pigs, or monkeys.

[0291] In some embodiments, the subject is a chicken, duck, goose, or bird.

[0292] In some embodiments, the subject cell, tissue or organ is from, but is not limited to, the eye, ear, nose, mouth, including gums and tooth roots; bone, lung, breast, pancreas, stomach, esophagus; muscle, including myocardium; liver, blood vessels; skin, including hair; heart, brain, nervous tissue, kidney, testicle, prostate, penis, cloaca, fin, ovary or intestine.

[0293] In some embodiments, the composition for rejuvenating cells, tissues, or organs of a subject is capable of restoring epigenetic information to the cells, tissues, or organs of the subject.

[0294] In some embodiments, the composition for rejuvenating cells, tissues, or organs in a subject is capable of restoring epigenetic information lost in the cells, tissues, or organs of the subject due to aging, injury, or disease.

[0295] In some embodiments, promoter expression can be used to reduce and reverse epigenetic marks associated with aging, increase epigenetic marks associated with cell rejuvenation, reduce the expression of aging-associated proteins, increase the expression of proteins associated with cell rejuvenation, restore the balance between euchromatin and heterochromatin, prevent the loss of cell identity, restore cell identity, reverse aging-associated DNA methylation changes, and thus rejuvenate cells.

[0296] In some embodiments, the SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC is a human protein, or a non-human protein, for example, a mammalian (e.g., mouse, rat, cow, sheep, horse, dog, cat, pig, monkey, chicken, duck, goose, bird) protein, or a protein that has at least 80% identity to the amino acid sequence of the corresponding human protein or non-human protein and maintains activity.

[0297] In some embodiments, the SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC or any combination thereof is a nucleic acid encoding a human protein, or a nucleic acid encoding a non-human protein, for example, a nucleic acid encoding a protein of a mammal (e.g., mouse, rat, cow, sheep, horse, dog, cat, pig, monkey, chicken, duck, goose, bird), or a nucleic acid that has at least 80% identity with a corresponding nucleic acid encoding a human protein, or a nucleic acid sequence encoding a non-human protein and retains activity.

[0298] In some embodiments, the method induces cell reprogramming, reverses aging, improves tissue function, improves organ function, promotes tissue repair, promotes tissue survival, promotes tissue regeneration, promotes tissue growth, promotes angiogenesis, reduces scar formation, alleviates external manifestations of aging including alopecia, hair thinning, hair graying, skin laxity and skin wrinkling, promotes organ regeneration, promotes organ survival, treats disease, or any combination thereof.

[0299] In some embodiments, the method reverses aging of a cell, tissue, organ, or subject, thereby rejuvenating the cell, tissue, or organ.

[0300] In some embodiments, the methods can promote neuronal regeneration in a subject.

[0301] In some embodiments, the composition can protect neurons in a subject from damage or degeneration.

[0302] In some embodiments, the method can reverse the epigenetic age of senescence in skin fibroblasts of a subject.

[0303] In some embodiments, the methods can reverse senescence of skin fibroblasts in a subject.

[0304] In some embodiments, the methods can reverse senescence of chondrocytes in a subject.

[0305] In some embodiments, the methods can reverse aging of muscle stem cells in a subject.

[0306] In some embodiments, the method can promote improvement of osteoarthritis symptoms in a subject.

[0307] In some embodiments, the methods can promote improvement of glaucoma symptoms in a subject.

[0308] In some embodiments, the composition can promote regeneration of the optic nerve.

[0309] In some embodiments, the composition can promote axonal regeneration in the optic nerve.

[0310] In some embodiments, the composition can inhibit apoptosis of optic nerve cells (eg, retinal ganglion cells) in optic nerve damage.

[0311] In some embodiments, the method does not induce teratoma formation, or does not induce tumor growth or tumor formation.

[0312] In some embodiments, the method does not induce complete reprogramming.

[0313] In some embodiments, the method does not reprogram the cell to a pluripotent state, or the method does not reprogram the cell to an "induced pluripotent stem cell" or "iPSC."

[0314] In a third aspect, the present invention provides use of the composition of the first aspect in the preparation of a medicament for rejuvenating cells, tissues or organs of a subject.

[0315] In some embodiments, rejuvenating a cell, tissue, or organ of a subject comprises restoring a transcriptional profile or epigenetic information lost in the at least one cell, tissue, or organ due to aging, injury, disease, or any combination thereof.

[0316] In some embodiments, rejuvenating the subject's cells, tissues, or organs comprises restoring the function of the cells, increasing the potency of the cells, enhancing the viability of the cells, or increasing the replicative capacity or lifespan of the cells, or a combination thereof.

[0317] In some embodiments, the rejuvenation of the subject's cells, tissues, or organs comprises promoting axon regeneration in the subject's neuronal cells; reversing the epigenetic age of senescence in the subject's skin fibroblasts; reversing the aging of the subject's skin fibroblasts; reversing the aging of the subject's chondrocytes; reversing the aging of the subject's muscle stem cells; promoting improvement in the subject's osteoarthritis symptoms; or promoting improvement in the subject's eye disease symptoms.

[0318] In a fourth aspect, the present invention provides a kit comprising an induction factor for restoring the viability of somatic cells in a subject, comprising the induction factor. In some embodiments of the present invention, the kit further comprises reagents and vectors for reprogramming cells. The vector may be, for example, a lentiviral vector. The kit of the present invention may also include culture medium and instructions.

[0319] In some embodiments of the present invention, the induction factors used include: any one selected from SOX1, SOX2, SOX3, RCOR2, GMNN, and GLIS1; any one selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, and PAX1; and Klf4 or Klf2. In some embodiments of the present invention, the kit of the present invention further includes c-MYC.

[0320] In some embodiments, the subject has, is suspected of having, or is at risk of having:

[0321] Diseases, conditions or manifestations associated with impaired cell, tissue or organ vitality or aging;

[0322] The disease, condition or manifestation associated with impaired cell, tissue or organ vitality or aging is a neurodegenerative disease, cardiovascular disease, metabolic disease, musculoskeletal disease, inflammatory disease, manifestation or condition associated with the skin, eye disease, fibrotic disease, or disease associated with the reproductive system;

[0323] The neurodegenerative disease is cerebral stroke, Huntington's disease, senile dementia, Alzheimer's disease, Parkinson's disease;

[0324] The cardiovascular disease is hypertension, coronary heart disease arrhythmia, heart disease, hypotension, heart failure, angina pectoris, arteriosclerosis, myocardial infarction, myocarditis, congestive heart failure, atrioventricular block, atherosclerosis, myocardial infarction;

[0325] The metabolic disease is type I diabetes, type II diabetes, hyperglycemia, hyperinsulinemia, insulin resistance, obesity, gout;

[0326] The musculoskeletal disease is muscular dystrophy, myasthenia gravis, osteoporosis, osteoarthritis, osteochondrosis, osteochondritis, osteonecrosis, and osteopenia;

[0327] The inflammatory disease is systemic lupus erythematosus, polymyalgia rheumatica, gouty arthritis, degenerative arthritis, rheumatoid arthritis, inflammatory arthritis, Hashimoto's thyroiditis, inflammatory bowel disease, hepatitis, pneumonia, respiratory tract inflammation, encephalitis;

[0328] The skin-related aging symptoms or diseases are wrinkles, age spots, seborrheic keratosis, tinea pedis, urticaria, neurodermatitis, and pigmented nevi;

[0329] The eye disease is macular degeneration, retinal degeneration, retinal detachment, diabetic retinopathy, glaucoma, optic atrophy, floaters, cataract, non-arteritic anterior ischemic optic neuropathy, chorioretinitis, retinitis pigmentosa, optic nerve damage;

[0330] The fibrotic disease is pulmonary fibrosis, liver fibrosis, myocardial fibrosis, renal interstitial fibrosis, myelofibrosis, pleural fibrosis, intestinal wall fibrosis;

[0331] Diseases related to the reproductive system include erectile dysfunction and premature ovarian failure.

[0332] The present invention screens a new combination of aging reprogramming factors, which will have broad application prospects in the treatment and / or improvement of many aging-related or cell viability-impaired diseases and disorders.

[0333] The present invention unexpectedly discovered that a three-factor combination of any one factor selected from SOX1, SOX2, SOX3, RCOR2, GMNN and GLIS1, any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4 and PAX1, and factor Klf4 or Klf2, or a combination of any one factor selected from SOX1, SOX2, SOX3, RCOR2, GMNN and GLIS1, any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4 and PAX1, factor Klf4 or Klf2, and factor c-MYC can achieve the reversal of senescent cells or the restoration of cell vitality.

[0334] In the present invention, a "factor" refers to a biologically active protein, or a nucleic acid encoding the same, such as DNA or RNA, or a mixture thereof, which acts on cells to alter transcription and, upon expression, converts senescent cells into young cells. In some embodiments, the factor can be non-integrating, i.e., provided to the recipient somatic cell in a form that does not result in integration of the exogenous DNA into the recipient cell's genome.

[0335] The factors in the present invention can be transcription factors, including but not limited to SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, and c-MYC.

[0336] The reprogramming in the present invention is incomplete reprogramming and / or partial reprogramming and / or transient reprogramming. It should be understood that the "incomplete" and / or "partial" and / or "transient" reprogramming mentioned herein is compared with cells with a high level of ability (e.g., ES cells or iPS cells). In another embodiment, the reprogramming of somatic cells is incomplete and / or partial and / or transient reprogramming compared to iPS cells. Wherein after administration of the composition, the rejuvenated cells, tissues or organs do not express at least one stem cell marker (e.g., NANOG). Molecular markers associated with partial reprogramming to restore a youthful state include: epigenetic age, transcriptome characteristics, number of γ-H2AX foci, concentration of reactive oxygen species, enrichment of histone marks (e.g., H3K9me3 and H4K20me3), collagen levels, vimentin and E-cadherin levels, aging-related β-galactosidase activity, cell proliferation rate and / or karyotype characteristics.

[0337] As used herein, "incomplete" and / or "partial" and / or "transient" reprogramming includes where the reprogrammed somatic cells retain and / or comprise the phenotype of an unreprogrammed somatic cell. Such retention and / or inclusion of the unreprogrammed somatic cell phenotype includes where the expression of surface markers indicative of the cell lineage or identity of the somatic cell is retained. In addition, such retention and / or inclusion may also include where epigenetic markers of the lineage or identity of the unreprogrammed somatic cell are retained and / or included by the reprogrammed somatic cell. Thus, in one embodiment, the reprogrammed somatic cell comprises the phenotype of an unreprogrammed somatic cell. In another embodiment, the reprogrammed somatic cell retains and / or comprises the phenotype of an unreprogrammed somatic cell of the tissue from which the reprogrammed somatic cell was obtained. In another embodiment, the reprogrammed somatic cell retains and / or comprises the phenotype and / or epigenetic markers indicative of the cell lineage or identity of the somatic cell.

[0338] Epigenetic changes are one of the most important characteristics of aging, which are mainly manifested in the changes in the methylation status of the whole genome at the chromatin level with age.

[0339] Measuring cell age in different tissue types using epigenetic age is used to predict aging and validate effective anti-aging interventions. Furthermore, perturbations in the autophagy-lysosome system in senescent cells can lead to an increase in lysosome content, resulting in the accumulation of β-galactosidase, a lysosomal senescence-associated enzyme, in senescent cells. Therefore, histochemical analysis of β-galactosidase levels in cells is also a common method for assessing cellular senescence.

[0340] As used herein, cells include any type of cell that makes up an organism. Thus, somatic cells may include, for example, cells of the eye, ear, nose, mouth, including gums and tooth roots, bone, cartilage, lung, breast, udder, pancreas, stomach, esophagus, muscle, including myocardium, liver, blood vessels, skin, including hair, heart, brain, nervous tissue, kidney, testicle, prostate, penis, cloaca, fin, ovary, or intestine. In one embodiment of the present invention, the somatic cell is a cell from connective tissue, such as a skin fibroblast. In one embodiment, the somatic cell is a chondrocyte. In one embodiment, the somatic cell is a muscle stem cell. In another embodiment, the somatic cell is a neural cell, such as a cell from the central and / or peripheral nervous system. Thus, in one embodiment, the cell is a visual neuron. In another embodiment, the cell is a sensory neuron. In an alternative embodiment, the cell is a motor neuron. In another embodiment, the cell is an interneuron. In one embodiment, the somatic cell is from an animal. In another embodiment, the somatic cell is from a mammal. In another embodiment, the mammal is human. Thus, in a specific embodiment, the somatic cell is from a human and is a human somatic cell. In an alternative embodiment, the mammal is a mouse and the somatic cell is a mouse somatic cell.

[0341] In another embodiment, the reprogrammed somatic cells comprise molecular markers, such as epigenetic markers, that indicate a younger or smaller epigenetic age than non-reprogrammed somatic cells or somatic cells from a tissue or organism from which the reprogrammed somatic cells were obtained. In other embodiments, the reprogrammed somatic cells produced according to the methods defined herein comprise phenotypes and / or molecular markers, such as epigenetic markers, that are similar to the phenotypes and / or molecular markers of non-reprogrammed somatic cells.

[0342] In some embodiments, the tissue or organ defined herein is selected from the group consisting of: eye, ear, nose, mouth, including gums and tooth roots, bone, cartilage, lung, breast, udder, pancreas, stomach, esophagus, muscle, including myocardium, liver, blood vessels, skin, including hair, heart, brain, nervous tissue, kidney, testicle, prostate, penis, cloaca, fin, ovary or intestine.

[0343] In some embodiments, the composition induces cell reprogramming, reverses aging, improves tissue function, improves organ function, promotes tissue repair, promotes tissue survival, promotes tissue regeneration, promotes tissue growth, promotes angiogenesis, reduces scar formation, alleviates external manifestations of aging including alopecia, hair thinning, hair graying, skin sagging and skin wrinkling, promotes organ regeneration, promotes organ survival, treats disease, or any combination thereof.

[0344] In some embodiments, the method does not induce teratoma formation or does not induce tumor growth or tumor formation.

[0345] Herein, induction factor is used in its broadest sense and includes nucleic acids, chemicals, engineered proteins, any recombinant viruses that produce reprogrammed cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0346] Figures 1A to 1L show that reprogramming restores the aging physiological properties in aging mouse skin fibroblasts to a younger state. Figures 1A and 1B show the expression of related genes in cells after lentiviral transfection. Figures 1C and 1D show representative graphs of changes in β-galactosidase activity before and after treatment: 5 days of reprogramming using GATA6+SOX3+Klf4 and NKX3.1+SOX3+KLF4 compositions. Figures 1E and 1F show changes in DNA methylation age (i.e., changes in epigenetic age) in mouse skin fibroblasts after reprogramming using GATA6+SOX3+Klf4 and NKX3.1+SOX3+KLF4 compositions, respectively. Figures 1G to 1L show the effects of different combinations of GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, SOX1, SOX2, SOX3, RCOR2, GMNN, Klf2, Klf4, GLIS1, and c-MYC genes on β-galactosidase activity. All bar graphs were generated after merging all single cell data from biological and technical replicates for ease of viewing. The significance levels shown allow for a leniency in pairwise comparisons of differences between patients.

[0347] Figures 2A to 2J show that reprogramming promotes neuronal survival and axonal regeneration in human SH-SY5Y neuronal cells after vincristine (VCR)-induced damage. Figures 2A and 2B show the expression of relevant genes in cells after lentiviral transfection. Figures 2C and 2D show representative graphs of changes in synaptic length before and after treatment: 6 days of reprogramming using GATA6+SOX3+Klf4 and NKX3.1+SOX3+KLF4 combinations. Figures 2E to 2J show the effects of different combinations of GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, SOX1, SOX2, SOX3, RCOR2, GMNN, Klf2, Klf4, GLIS1, and c-MYC genes on synaptic length. All bar graphs were generated after merging all single cell data from biological and technical replicates for ease of viewing. The significance levels shown allow for one pairwise comparison of differences between patients.

[0348] Figures 3A to 3F show that reprogramming restores aging physiological properties in chondrocytes to a younger state: β-galactosidase activity in chondrocytes was measured after 5 days of reprogramming using different combinations of GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, SOX1, SOX2, SOX3, RCOR2, GMNN, Klf4, Klf2, GLIS1, and c-MYC genes. All bar graphs were generated after merging all single cell data from biological and technical replicates for ease of viewing. The significance level shown allows for a leniency in one pairwise comparison of differences between patients.

[0349] Figures 4A to 4F show that reprogramming restores the aging physiological properties of old mouse muscle stem cells to a younger state: β-galactosidase activity in mouse muscle stem cells was detected after 5 days of reprogramming using different combinations of GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, SOX1, SOX2, SOX3, RCOR2, GMNN, Klf4, Klf2, GLIS1, and c-MYC genes. All bar graphs were generated after merging all single cell data from biological and technical replicates for ease of viewing. The significance level shown allows for a leniency in one pairwise comparison of differences between patients.

[0350] Figure 5 shows the effect of reprogramming on promoting optic nerve regeneration. Figure 5A shows a schematic diagram of the ONC efficacy assay process; Figure 5B shows the AAV viral expression pattern; Figure 5C shows the effect of G2S1K2 on RGC apoptosis; Figure 5D shows the effect of G2S1K2 on optic nerve regeneration. DETAILED DESCRIPTION

[0351] The present invention is further illustrated by the following examples. Those skilled in the art will appreciate that the present invention is not limited to the examples described, and that those skilled in the art may modify the examples based on the teachings of the specification. Such modifications are also within the scope of the present invention as defined by the appended claims.

[0352] The experimental methods in the following examples are conventional methods unless otherwise specified.

[0353] 1. Mouse strain

[0354] ICR mice were purchased from Vital River.

[0355] 2. Viral transfection

[0356] The coding nucleic acid sequences of the target factors were respectively inserted into the FUW-TetON lentiviral vector and transduced into 293T cells to package the lentivirus. After 72 hours, the viral supernatants were collected, passed through a 0.22μm membrane, and aliquoted and frozen at -80°C. In the presence of polybrene (10μg / ml), the supernatants containing the lentivirus expressing each target factor were mixed separately to obtain a lentiviral combination of the target factor, which was then used to simultaneously infect mouse skin fibroblasts, mouse chondrocytes, and mouse muscle stem cells. For the transfection of SH-SY5Y neuronal cells, it was performed under conditions without polybrene. 12 hours after transfection, the cells were replaced with a culture medium containing 1μg / ml doxycycline to induce expression for at least 3 days.

[0357] The induction factors used in the examples include:

[0358] GATA1(AAV38577.1), GATA2(NCBI Reference Sequence: NP_001139133.1), GATA3(AAX36808.1), GATA4(NCBI Reference Sequence:NP_001295022.1), GATA6(NCBI Reference Sequence:NP_005248.2), SOX1(NCBI Reference Sequence:NP_005977.2), SOX2(CAA83435.1), SOX3(NCBI Reference Sequence:NP_005625.2), SOX7(AAP36695.1), SOX17(BAB83867.1), OCT4(PDB:6T90_K), FOXA2(NCBI Reference Sequence:NP_068556.2), PAX1 (NCBI Reference Sequence:NP_006183.2), C / EBPα (NCBI Reference Sequence:NP_004355.2), NKX3.1 (NCBI Reference Sequence:NP_006158.2), NR5A2 (NCBI Reference Sequence:NP_995582.1), KLF2 (NCBI Reference Sequence:NP_057354.1), KLF4 (NCBI Reference Sequence:NP_004226.3), GMNN (NCBI Reference Sequence:NP_001238918.1), RCOR2 (NCBI Reference Sequence:XP_047282784.1), c-MYC (BAA01374.2) and GLIS1 (NCBI Reference Sequence:NP_001354413.1).

[0359] 3. Extraction of skin fibroblasts from aged mice

[0360] 24-month-old mice were euthanized and their dorsal skin was shaved and excised. Before excision, the skin was cleaned with povidone-iodine and rinsed with cold 1× PBS. Skin fibroblasts were isolated using trypsin and collagenase. Fibroblasts were resuspended and cultured in DMEM GlutaMax medium.

[0361] 8-week-old mice were selected as young mice.

[0362] 4. Differentiation of SH-SY5Y cells into neurons

[0363] One day after plating, cells began to differentiate in EMEM / F12 medium (1:1) containing 5% FBS and 10 μM all-trans retinoic acid (RA, MCE) (differentiation medium 1) for 5 days (medium was changed every three days), and then the medium was changed to serum-free neurobasal / B27 plus medium (Gibco) containing 1× Glutamax (Gibco) and 50 ng / ml BDNF (MCE) (differentiation medium 2) for maturation for 5 days.

[0364] 5. Induction of chondrocyte senescence

[0365] Human chondrocyte cell line C28 / I2 was plated. One day after plating, 20 μM etoposide was added to the cells for senescence induction. 24 hours after induction, the culture medium was replaced with fresh complete medium DMEM (Gibco) (containing 10% FBS (Gibco) and 1% P / S (Gibco)) and cultured for a further 3 days. The composition was then added for treatment for 5 days before fixation and subsequent treatment.

[0366] 6. Extraction of muscle stem cells from aged mice

[0367] Fifteen-month-old mice were selected and sacrificed by cervical dislocation, and the muscles were separated, cut into small pieces, and incubated in DMEM medium (DMEM; ThermoFisher Scientific) containing 0.2% type I collagenase (Sigma-Aldrich) at 37°C for 2 hours. Single fibers were collected using a pipette tip and purified twice in DMEM containing 1 g / L glucose (supplemented with 10% horse serum (HS, ThermoFisher Scientific), 1% penicillin / streptomycin (PS, ThermoFisher Scientific), and 0.5% chick embryo extract (CEE, ThermoFisher Scientific). The single fibers were then transferred and the myofiber suspension was passed through a syringe with a 21G needle through a 40 μm filter. The harvested stem cells were directly plated onto culture dishes coated with Matrigel growth factor (GFR) basement membrane matrix (Corning) or onto culture dishes containing coverslips coated with Matrigel GFR basement membrane matrix. The cells were expanded in DMEM containing 1 g / L glucose supplemented with 10% HS, 20% FBS, 1% PS, and 0.5% CEE and cultured at 37°C, 5% CO2. The culture medium was changed every 2 days.

[0368] 7. β-galactosidase activity detection

[0369] After treatment, cells were fixed with 4% paraformaldehyde for 15 minutes and then stained in situ using a β-galactosidase in situ staining kit (Beyotime). Following staining, DAPI was added to stain the nuclei for cell count. Cells were then observed under a fluorescence microscope, and the percentage of positive cells was calculated (positive cell percentage = number of positively stained cells / total number of cells × 100%).

[0370] 8. Epigenetic age testing

[0371] The genome was extracted and purified using a DNA extraction kit (Qiagen), and the methylation sites in the target gene region were sequenced. The cell's age was calculated based on the sequencing results of the target CpG sites.

[0372] 9. Neurite Regeneration Assay

[0373] Mature neurons differentiated from SH-SY5Y cells were transduced with a lentiviral vector. Following transduction, the cells were cultured for at least three days before inducing neurite regeneration by adding 200 nM vincristine (VCR, MCE) for 24 hours. Following vincristine treatment, the neurons were washed twice in PBS, and fresh differentiation medium 2 was added back to the plate for an additional two weeks to monitor neurite regeneration.

[0374] 10. Immunofluorescence and Imaging

[0375] After fixation with 4% paraformaldehyde for 30 minutes at room temperature, cells were permeabilized and blocked with 1% BSA containing 0.25% Triton X-100 for 1 hour. Cells were incubated with appropriate ratios of primary antibodies in 1% BSA containing 0.25% Triton X-100 overnight at 4°C. After washing with 0.1% BSA, cells were incubated with secondary antibodies for 1 hour at room temperature. All secondary antibodies were Alexa Fluor-conjugated (Molecular Probes) and diluted 1:1000 in 1% BSA. After washing again with 0.1% BSA, cell nuclei were counterstained with 5 μg / mL LDAPI in PBS for 5 minutes. After washing with PBS, optical sections were captured using a fluorescence microscope. Semi-quantitative fluorescence analysis was performed using ImageJ.

[0376] Example 1. Reversal of cellular senescence by reprogramming

[0377] To investigate the possibility of reprogramming to reverse aging, we infected skin fibroblasts from aged mice with lentiviral vectors encoding human GATA6, SOX3, NKX3.1, and KLF4 under a Tet-inducible promoter (using the Tet-on system) in different combinations as shown below.

[0378] FIG1A shows that under G6S3K4 On condition, the expression of GATA6, SOX3, and KLF4 was induced in cells.

[0379] FIG. 1B shows that under G6S3K4 On conditions, the expression of NKX3.1, SOX3, and KLF4 was induced in cells.

[0380] After infection, we treated the cells with doxycycline for 5 days and then examined changes in cellular senescence using β-galactosidase activity assays and epigenetic age analysis. β-galactosidase is one of the earliest and most widely used lysosomal senescence markers. It increases lysosomal biogenesis in senescent cells and is considered a specific marker of cellular senescence and a gold standard for senescence detection.

[0381] β-galactosidase levels in skin fibroblasts from aged mice were measured 5 days after doxycycline induction. Compared to untreated cells (Aged group), cells in which GATA6, SOX3, and Klf4 expression was induced (G6S3K4On condition) showed decreased β-galactosidase activity (Figure 1C). Cells in which NKX3.1, SOX3, and KLF4 expression was induced (G6S3K4On condition) also showed decreased β-galactosidase activity (Figure 1D).

[0382] Similarly, epigenetic age analysis results also showed that compared with untreated cells (Aged group), cells in which GATA6, SOX3, and Klf4 expression was induced (G6S3K4 On condition) (Figure 1E) showed a significant reversal effect of methylation age, which could reverse cell age by approximately 8 months. Cells in which NKX3.1, SOX3, and KLF4 expression was induced (Figure 1F) (G6S3K4 On condition) also showed a significant reversal effect of methylation age, which could reverse cell age by approximately 6 months.

[0383] Furthermore, lentiviral vectors expressing GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, SOX1, SOX2, SOX3, RCOR2, GMNN, KLF2, KLF4, GLIS1, and c-MYC were combined to infect cells, and then the activity of β-galactosidase in skin fibroblasts of aged mice was examined. The results showed that the expression of the above gene combinations, such as NR5A2+SOX2+KLF4, FOXA2+SOX1+KLF4, OCT4+RCOR2+Klf4+c-MYC, and FOXA2+SOX1+KLF4+c-MYC, significantly reduced the activity of β-galactosidase in cells (Figure 1G to Figure 1L).

[0384] Example 2. Reversal effect of reprogramming on SH-SY5Y neuronal cell damage

[0385] To determine whether expression of human GATA6, human SOX3, and human Klf4 (human G6S3K4), as well as human NKX3.1, human SOX3, and human KLF4 (human N3S3K4), could protect human neuronal cells and regenerate neuronal synapses in vitro, a neurite regeneration assay was used as described below. SH-SY5Y cells are human neuroblastoma cells that can differentiate into neurons and were transduced with lentiviral vectors encoding human GATA6, human SOX3, and human Klf4 (tetOn-hG6S3K4), encoding human NKX3.1, human SOX3, and human KLF4 (tetOn-h N3S3K4), or with a lentiviral vector encoding mCherry under a Tet-inducible promoter (using the Tet-on system).

[0386] FIG2A shows that under the G6S3K4 On condition, the expression of human GATA6, human SOX3, and human Klf4 was induced in cells.

[0387] FIG2B shows that under the N3S3K4 On condition, the expression of human NKX3.1, human SOX3, and human KLF4 was induced in cells.

[0388] 3 days after transduction, cells were treated with vincristine (VCR) for 24 hours to induce neurite degeneration. VCR is a chemotherapy drug that destroys microtubules. It is typically used in vitro to determine whether damage post-treatment maintains and / or promotes cell function (e.g., neuronal function). As described herein, VCR was used to determine the effect of G6S3K4 treatment on neuronal survival and synaptic regrowth. After VCR treatment, cells were grown in differentiation medium and the length of neurites was measured.

[0389] Cell was processed with VCR and the neuron length of cell was measured in 9 days after 24 hours.Relative to the cell (mCherry On condition) for wherein inducing mCherry expression, the cell (G6S3K4On condition) for wherein inducing human GATA6, human SOX3 and human Klf4 expression showed increased neuron survival and synaptic length (Fig. 2 C, left figure).Similarly, the quantitative display of neurite length was compared with mCherry expression, and human GATA6, human SOX3 and human Klf4 expression increased neurite length by at least 4 times (Fig. 2 C, right figure).

[0390] Figure 2D (left panel) shows that cells with induced expression of human NKX3.1, human SOX3, and human KLF4 (N3S3K4 On condition) show increased neuronal survival and axon length. Figure 2D (right panel) shows that quantification of neurite length shows that expression of human NKX3.1, human SOX3, and human KLF4 increased neurite length by at least 3-fold compared to mCherry expression.

[0391] Furthermore, different combinations of GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, SOX1, SOX2, SOX3, RCOR2, GMNN, KLF2, KLF4, GLIS1, and c-MYC genes were expressed in SH-SY5Y cells, and the neurite length was investigated to detect the effects of these combinations on neurite regeneration.

[0392] The results showed that the expression of the above combinations, such as NR5A2+SOX2+Klf4, OCT4+SOX3+Klf4+c-MYC, FOXA2+SOX3+KLF4, NKX3.1+SOX1+KLF2+c-MYC, etc., can significantly promote neurite regeneration (Figure 2E to Figure 2J).

[0393] These results indicate that expression of human G6S3K4, N3S3K4 and other combinations protects human neuronal cells from VCR-induced neuronal degeneration.

[0394] Example 3. Reversal of chondrocyte senescence by reprogramming

[0395] To determine the reversal effect of the new reprogramming combination on chondrocyte aging, a β-galactosidase activity assay was used as described below. Different combinations of GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, SOX1, SOX2, SOX3, RCOR2, GMNN, KLF4, KLF2, GLIS1, and c-MYC genes were expressed in senescent human chondrocytes. After 5 days of reprogramming, β-galactosidase activity in the chondrocytes was measured. The results showed that the new gene combination could reduce β-galactosidase activity in the cells by 60% to 70% compared to untreated cells (Aged group) (Figures 3A to 3F).

[0396] Example 4. Reversal of aging effects of reprogramming on muscle stem cells in aged mice

[0397] To determine the aging-reversing effect of the new reprogramming combination on aged mouse muscle stem cells, a β-galactosidase activity assay was used as described below. Different combinations of GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, or PAX1, SOX1, SOX2, SOX3, RCOR2, GMNN, KLF4, KLF2, GLIS1, and c-MYC genes were expressed in aged mouse muscle stem cells. After 5 days of reprogramming, β-galactosidase activity in aged mouse muscle stem cells was measured. The results showed that the new gene combination could reduce β-galactosidase activity in cells by approximately 60% to 70% compared to untreated cells (Aged group) (Figures 4A to 4F).

[0398] Example 5. Effect of reprogramming on promoting optic nerve regeneration

[0399] To further test whether the G2S1K2 (GATA2+SOX1+KLF2) combination can significantly promote optic nerve regeneration in mice and exert an aging reprogramming effect in vivo, this example uses a mouse optic nerve injury model for in vivo efficacy testing. Figure 5A illustrates the schematic flow chart of the in vivo efficacy experiment. The specific method is as follows:

[0400] 1. Preparation of AAV-TRE-G2S1K2 and AAV-CAG-tTA viruses:

[0401] The coding nucleic acid sequences of GATA2, SOX1, and KLF2 were concatenated together through linkers P2A and T2A, respectively (GATA2-P2A-SOX1-T2A-KLF2) and inserted into the AAV-TRE vector (containing the tetracycline response element (TRE3G promoter) and the SV40 element) to form the AAV-TRE-G2S1K2 vector ( Figure 5B ). AAV-CAG-tTA was purchased from Wuhan Shumi Brain Science Technology Co., Ltd.

[0402] 2. Construction of Optic Nerve Crushing Model:

[0403] Approximately 4-week-old mice were anesthetized and intravitreally injected with 1.5 μl of viral solution, consisting of 1 μl of AAV-TRE-G2S1K2 and 0.5 μl of AAV-CAG-rTA. Following viral injection, the mice were divided into G2S1K2 and G2S1K2+dox groups, with 5 mice in each group. The G2S1K2+dox group received 2 mg / ml of doxycycline hydrochloride (Dalian Meilun Biotechnology, #MB1088-2) in their drinking water to inhibit G2S1K2 expression in the virus, while the G2S1K2 group received normal drinking water.

[0404] Two weeks after AAV injection, mice were deeply anesthetized and, under a dissecting microscope, the optic nerve was pinched with scissors approximately 2 mm from the eyeball for approximately 5 seconds. The forceps were then withdrawn. Two weeks later, cholera toxin B subunit (CTB) (thermo, #C34775) was injected intravitreally to fluorescently label the optic nerve. Two days later, the mice were euthanized and eye samples were obtained.

[0405] 3. Eyes:

[0406] Mice were euthanized using CO2, and their eyeballs and optic nerves were removed.

[0407] 4. Retinal flat mount:

[0408] After the removal of the eyeball, it was rinsed with PBS and placed in 4% paraformaldehyde solution and fixed at room temperature for 1.5-2 hours. The anterior cornea, lens and vitreous body of the fixed mouse eyeball were removed in PBS, and the retina was divided into 4 petals. 0.5% Triton X-100 was added and the membrane was permeabilized on a horizontal shaker for 30 minutes. The 0.5% Triton X-100 was aspirated and blocking solution (5% BSA solution containing 0.1% Triton X-100 prepared in PBS) was added and blocked overnight at 4°C on a horizontal shaker. RBPMS antibody working solution (RBPMS, Abcam, #AB152101, 1:300 dilution ratio) was added and incubated in a 4°C refrigerator for 2 days. PBS was washed 3 times on a shaker. Alexa Fluor TMThe cells were incubated with 488 donkey anti-rabbit IgG working solution (Thermo Fisher, #A-21206, 1:1000 dilution) on a shaker at room temperature for 2.5 hours. The retinas were aspirated and mounted on coverslips. Images were taken using a confocal microscope at 20× magnification. The number of RBPMS-positive cells (i.e., retinal ganglion cells (RGCs)) in each image was counted using Image software. The results are shown in Figure 5C.

[0409] 5. Optic nerve frozen section:

[0410] After removal, the eyeballs were rinsed with PBS and fixed overnight in paraformaldehyde. The optic nerves were then placed in a 30% sucrose solution and dehydrated at 4°C for approximately 1.5 days. The optic nerves were embedded in OCT embedding medium and fixed in a -80°C freezer. Frozen sections were made at -20°C, with a thickness of 8-10 μm. After washing the OCT off the slides with PBS, anti-fluorescence attenuation mounting medium was added, and a coverslip was applied. The entire optic nerve was photographed using a confocal microscope at 20× magnification.

[0411] Results: Image J software was used for analysis. A starting point was selected and the number of axons at that distance was calculated from the starting point in the direction of weak fluorescence. The results are shown in Figure 5D.

[0412] The results showed that compared with the G2S1K2+dox group, the G2S1K2 group could significantly inhibit RGC cell apoptosis caused by crush injury (Figure 5C) and significantly promote axon regeneration in the optic nerve (Figure 5D), indicating that G2S1K2 has an aging reprogramming effect.

Claims

1. A composition for rejuvenating cells, tissues or organs of a subject, the composition comprising: i) any one factor selected from SOX1, SOX2, SOX3, RCOR2, GMNN and GLIS1, ii) any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4 and PAX1, and iii) Factor Klf4 or Klf2.

2. The composition according to claim 1, characterized in that The composition also includes a c-MYC factor.

3. The composition according to claim 2, characterized in that The factors SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC are proteins, nucleic acids encoding them, or mixtures thereof, Preferably, the factors SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC are proteins, fusion proteins bound to the membrane permeability domain, and are in the form of nucleic acids encoding them.

4. The composition according to any one of claims 1 to 3, characterized in that The factors SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC are in the form of expression vectors, and the expression vectors are plasmid vectors, episomal vectors, transposons, or viral vectors, Preferably, the viral vector is an adenoviral vector, an adeno-associated viral vector (AAV), a lentiviral vector, a Sendai viral vector, a cytomegalovirus vector or a vaccinia virus vector. Preferably, the factors are present on the same expression vector, or on multiple expression vectors.

5. The composition according to any one of claims 1 to 3, characterized in that The factors SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC are in the form of DNA or RNA, Preferably, the factor is in the form of mRNA, The factors are connected in series on the same mRNA molecule, or the factors SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC are in the form of a complex of different mRNA molecules, Preferably, the factor is a synthetic mRNA, which encodes a wild-type form, a mutant, or a genetically engineered form of a gene selected from the group consisting of SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, and c-MYC.

6. The composition according to any one of claims 1 to 3, characterized in that The subject is a mammal, including but not limited to humans, mice, rats, cattle, sheep, horses, dogs, cats, pigs or monkeys, The subject's cells, tissues or organs are from, but not limited to, eyes, ears, nose, mouth, including gums and tooth roots; Bones, lungs, breasts, pancreas, stomach, esophagus; muscles, including myocardium; liver, blood vessels; Skin, including hair; heart, brain, nervous tissue, kidneys, testicles, prostate, penis, cloaca, fins, ovaries or intestines.

7. A composition according to any one of claims 1 to 3, characterized in that The factors SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC are human proteins, or non-human proteins, or proteins that have at least 80% identity with the amino acid sequence of the human proteins or non-human proteins and maintain activity, The non-human protein is a corresponding protein of mouse, rat, cattle, sheep, horse, dog, cat, pig, monkey, chicken, duck, goose or bird; or The factors SOX1, SOX2, SOX3, RCOR2, GMNN, GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4, PAX1, Klf4, Klf2, GLIS1, c-MYC are nucleic acids encoding human proteins, or nucleic acids encoding non-human proteins, or nucleic acids having at least 80% identity with corresponding nucleic acids encoding human proteins, or nucleic acids encoding non-human proteins and maintaining activity, The nucleic acid is a nucleic acid encoding the corresponding protein of mouse, rat, cattle, sheep, horse, dog, cat, pig, monkey, chicken, duck, goose, and bird.

8. A method for rejuvenating cells, tissues or organs of a subject, characterized in that: The method comprises administering to a subject a composition according to any one of claims 1 to 7.

9. Use of the composition of any one of claims 1 to 7 in the preparation of a medicament for rejuvenating cells, tissues or organs of a subject.

10. The method according to claim 8 or the use according to claim 9, characterized in that The rejuvenating the subject's cells, tissues or organs includes restoring the transcriptional profile or epigenetic information lost in at least one of the cells, tissues or organs due to aging, injury, disease or any combination thereof.

11. The method according to claim 8 or the use according to claim 9, characterized in that The rejuvenating the subject's cells, tissues or organs includes restoring the function of the cells, increasing the potential of the cells, enhancing the survival ability of the cells, or increasing the replication ability or lifespan of the cells, or a combination thereof.

12. The method according to claim 8 or the use according to claim 9, characterized in that The rejuvenation of the subject's cells, tissues or organs includes promoting axon regeneration in the subject's neuronal cells; reversing the epigenetic age of aging in the subject's skin fibroblasts; reversing the aging of the subject's skin fibroblasts; reversing the aging of the subject's chondrocytes; reversing the aging of the subject's muscle stem cells; promoting improvement of the subject's osteoarthritis symptoms; or promoting improvement of the subject's eye disease symptoms.

13. A kit comprising induction factors for restoring the viability of somatic cells of a subject, characterized in that: The induction factor comprises: i) any one factor selected from SOX1, SOX2, SOX3, RCOR2, GMNN and GLIS1, ii) any one factor selected from GATA1, GATA2, GATA3, GATA4, GATA6, FOXA2, SOX7, SOX17, NKX3.1, NR5A2, C / EBPα, OCT4 and PAX1, and iii) Factor Klf4 or Klf2.

14. The kit according to claim 13, characterized in that Inducible factors also include c-MYC.

15. The composition of any one of claims 1 to 7, the method of claim 8, the use of claim 9, the method or use of any one of claims 10 to 12, or the kit of any one of claims 13 to 14, characterized in that The subject has, is suspected of having, or is at risk of: Diseases, conditions or manifestations associated with impaired cell, tissue or organ viability or aging; The disease, condition or manifestation associated with impaired cell, tissue or organ vitality or aging is a neurodegenerative disease, cardiovascular disease, metabolic disease, musculoskeletal disease, inflammatory disease, manifestation or disorder associated with the skin, eye disease, fibrotic disease, or disease associated with the reproductive system; Preferably, the neurodegenerative disease is cerebral stroke, Huntington's disease, senile dementia, Alzheimer's disease, Parkinson's disease; Preferably, the cardiovascular disease is hypertension, arrhythmia of coronary heart disease, heart disease, hypotension, heart failure, angina pectoris, arteriosclerosis, myocardial infarction, myocarditis, congestive heart failure, atrioventricular block, atherosclerosis, myocardial infarction; Preferably, the metabolic disease is type I diabetes, type II diabetes, hyperglycemia, hyperinsulinemia, insulin resistance, obesity, gout; Preferably, the musculoskeletal disease is muscular dystrophy, myasthenia gravis, osteoporosis, osteoarthritis, osteochondrosis, osteochondritis, osteonecrosis, osteopenia; Preferably, the inflammatory disease is systemic lupus erythematosus, polymyalgia rheumatica, gouty arthritis, degenerative arthritis, rheumatoid arthritis, inflammatory arthritis, Hashimoto's thyroiditis, inflammatory bowel disease, hepatitis, pneumonia, respiratory tract inflammation, encephalitis; Preferably, the skin-related aging symptoms or diseases are wrinkles, age spots, seborrheic keratosis, tinea pedis, urticaria, neurodermatitis, and pigmented nevus; Preferably, the eye disease is macular degeneration, retinal degeneration, retinal detachment, diabetic retinopathy, glaucoma, optic atrophy, floaters, cataract, non-arteritic anterior ischemic optic neuropathy, chorioretinitis, retinitis pigmentosa, optic nerve damage; Preferably, the fibrotic disease is pulmonary fibrosis, liver fibrosis, myocardial fibrosis, renal interstitial fibrosis, myelofibrosis, pleural fibrosis, intestinal wall fibrosis; Preferably, the diseases related to the reproductive system are erectile dysfunction and premature ovarian failure.

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