Chemical reprogramming to generate neuronal cells
Inventors
Zhang, Mingliang • Ding, Sheng
Assignees
Publication Number
US-12104169-B2
Publication Date
2024-10-01
Expiration Date
2036-08-03
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Abstract
Compositions and methods are described herein for chemically inducing cells to change their differentiation state and become neuronal cells.
Core Innovation
The invention describes compositions and methods for chemically inducing differentiated, non-neuronal cells to change their differentiation state and become neuronal progenitor or mature neuronal cells without the need for genetic engineering. A key aspect is the use of a chemical cocktail comprising at least four active agents such as a BMP type I receptor ALK2/3 inhibitor, a TGF-beta inhibitor, a WNT inhibitor, neuronal differentiation enhancers, SMO agonists, retinoic acid receptor γ agonists, DNA methyltransferase inhibitors, histone demethylase inhibitors, and autophagy regulators to reprogram cells chemically.
The compositions and methods overcome the problem identified in the background that differentiated cell states are stable and resistant to lineage changes, and current genetic reprogramming methods pose risks of mutations due to insertion of expression cassettes. Furthermore, conventional methods using transcription factors have low efficiency and are time-consuming. Unlike prior approaches, this invention uses small molecules to modulate endogenous signaling pathways and epigenetic states to achieve neural fate reprogramming in a more efficient and less laborious manner.
The invention also addresses the challenge of unambiguous lineage tracing by using genetically marked fibroblasts to confirm that reprogrammed cells derive from differentiated fibroblasts, thereby demonstrating true chemical reprogramming rather than differentiation from contaminating precursor cells. The methods support the generation of expandable and tripotent neural stem-like cells capable of differentiating into neurons, astrocytes, and oligodendrocytes both in vitro and in vivo.
Claims Coverage
The independent claims cover methods and compositions related to chemical reprogramming of differentiated non-neuronal cells into neuronal cells using a specific cocktail of active agents.
Chemical composition for neuronal cell reprogramming
A composition comprising at least four active agents selected from BMP type I receptor ALK2/3 inhibitors, TGF-beta inhibitors, WNT inhibitors, neuronal differentiation enhancers, SMO agonists, retinoic acid receptor γ agonists, DNA methyltransferase inhibitors, histone demethylase inhibitors, and autophagy regulators, specifically including agents CHIR99021, LDN193189, A83-01, Hh-Ag1.5, retinoic acid, SMER28, RG108, parnate, and bFGF.
Method of generating neuronal or neuronal progenitor cells
A method comprising contacting a selected differentiated, non-neuronal cell with the nine-agent chemical composition (CHIR99021, LDN193189, A83-01, Hh-Ag1.5, retinoic acid, SMER28, RG108, parnate, and bFGF) in amounts sufficient to reprogram the cell to generate neuronal progenitor or mature neuronal cells.
Administration to subjects with neuronal diseases
Methods comprising administering the reprogrammed neuronal or progenitor cells, or the chemical composition, to subjects suffering from neuronal diseases or conditions, including but not limited to ALS, Alzheimer's disease, Parkinson's disease, multiple sclerosis, spinal muscular atrophy, and spinal cord injury.
The claims focus on the inventive chemical cocktail and its use to effectively reprogram differentiated non-neuronal cells to neuronal lineage cells without genetic manipulation, and on therapeutic methods administering these cells or compositions to treat neuronal diseases.
Stated Advantages
Avoids genetic engineering and associated risks of insertional mutations.
Provides a less labor-intensive, less time-consuming approach compared to transcription factor-based reprogramming.
Achieves higher efficiency reprogramming of differentiated fibroblasts to neuronal progenitor and mature neuronal cells.
Allows generation of expandable, tripotent neural stem cell-like cells from fibroblasts chemically.
Enables in vivo engraftment and differentiation of reprogrammed cells into functional neuronal, astrocyte, and oligodendrocyte lineages.
Documented Applications
Treatment of neurodegenerative diseases such as Amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Parkinson's disease, multiple sclerosis, Huntington's disease, and hereditary spastic paraplegia.
Therapeutic use in motor neuron diseases including Primary lateral sclerosis, Progressive bulbar palsy, Pseudobulbar palsy, and Spinal muscular atrophy (various types).
Treatment of traumatic injuries including stroke, head trauma, and spinal cord injury.
In vitro generation of neuronal progenitor and mature neuronal cells for transplantation.
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