RESUMEN
Astrocytes, due to the proximity to neuronal lineage and capability to proliferate, are ideal starting cells to regenerate neurons. Human fetal astrocytes have been successfully converted into neuronal cells by small molecules, which offered a broader range of further applications than transcription factor-mediated neuronal reprogramming. Here we report that human adult astrocytes could also be converted into neuronal cells by a different set of small molecules. These induced cells exhibited typical neuronal morphologies, expressed neuronal markers, and displayed neuronal electrophysiological properties. Genome-wide RNA-sequencing analysis showed that the global gene expression profile of induced neuronal cells resembled that of human embryonic stem cell-differentiated neurons. When transplanted into post-natal mouse brains, these induced neuronal cells could survive and become electrophysiologically mature. Altogether, our study provides a strategy to directly generate transgene-free neuronal cells from human adult astrocytes by small molecules.
Asunto(s)
Astrocitos/citología , Astrocitos/efectos de los fármacos , Transdiferenciación Celular/efectos de los fármacos , Reprogramación Celular/efectos de los fármacos , Neuronas/citología , Neuronas/efectos de los fármacos , Adulto , Animales , Astrocitos/metabolismo , Biomarcadores , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Supervivencia Celular/genética , Reprogramación Celular/genética , Análisis por Conglomerados , Fenómenos Electrofisiológicos , Células Madre Embrionarias/citología , Células Madre Embrionarias/metabolismo , Perfilación de la Expresión Génica , Humanos , Ratones , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Neuronas/metabolismo , TranscriptomaRESUMEN
Transplantation of embryonic γ-aminobutyric acid (GABA)ergic neurons has been shown to modify disease phenotypes in rodent models of neurologic and psychiatric disorders. However, whether transplanted interneurons modulate fear memory remains largely unclear. Here, we report that transplantation of embryonic interneurons into the amygdala does not alter host fear memory formation. Yet approximately 2 weeks after transplantation, but not earlier or later, extinction training produces a marked reduction in spontaneous recovery and renewal of fear response. Further analyses reveal that transplanted interneurons robustly form functional synapses with neurons of the host amygdala and exhibit similar developmental maturation in electrophysiological properties as native amygdala interneurons. Importantly, transplanted immature interneurons reduce the expression of perineuronal nets, promote long-term synaptic plasticity, and modulate both excitatory and inhibitory synaptic transmissions of the host circuits. Our findings demonstrate that transplanted immature interneurons modify amygdala circuitry and suggest a previously unknown strategy for the prevention of extinction-resistant pathological fear.
Asunto(s)
Amígdala del Cerebelo/fisiología , Extinción Psicológica/fisiología , Miedo/fisiología , Interneuronas/trasplante , Memoria/fisiología , Amígdala del Cerebelo/citología , Amígdala del Cerebelo/metabolismo , Animales , Conducta Animal , Condicionamiento Clásico/fisiología , Inmunohistoquímica , Interneuronas/metabolismo , Ratones , Inhibición Neural/fisiología , Plasticidad Neuronal , Técnicas de Placa-ClampAsunto(s)
Astrocitos/citología , Expresión Génica/efectos de los fármacos , Animales , Anticonvulsivantes/farmacología , Astrocitos/efectos de los fármacos , Glucógeno Sintasa Quinasa 3/antagonistas & inhibidores , Humanos , Ratones , Neuronas/citología , Pirazoles/farmacología , Piridinas/farmacología , Pirimidinas/farmacología , Factor de Crecimiento Transformador beta/antagonistas & inhibidores , Ácido Valproico/farmacologíaRESUMEN
Neuronal conversion from human fibroblasts can be induced by lineage-specific transcription factors; however, the introduction of ectopic genes limits the therapeutic applications of such induced neurons (iNs). Here, we report that human fibroblasts can be directly converted into neuronal cells by a chemical cocktail of seven small molecules, bypassing a neural progenitor stage. These human chemical-induced neuronal cells (hciNs) resembled hiPSC-derived neurons and human iNs (hiNs) with respect to morphology, gene expression profiles, and electrophysiological properties. This approach was further applied to generate hciNs from familial Alzheimer's disease patients. Taken together, our transgene-free and chemical-only approach for direct reprogramming of human fibroblasts into neurons provides an alternative strategy for modeling neurological diseases and for regenerative medicine.
Asunto(s)
Enfermedad de Alzheimer/patología , Fibroblastos/patología , Neuronas/citología , Bibliotecas de Moléculas Pequeñas/farmacología , Adulto , Fenómenos Electrofisiológicos/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Perfilación de la Expresión Génica , Humanos , Masculino , Neuronas/efectos de los fármacosRESUMEN
Neural progenitor cells (NPCs) can be induced from somatic cells by defined factors. Here we report that NPCs can be generated from mouse embryonic fibroblasts by a chemical cocktail, namely VCR (V, VPA, an inhibitor of HDACs; C, CHIR99021, an inhibitor of GSK-3 kinases and R, Repsox, an inhibitor of TGF-ß pathways), under a physiological hypoxic condition. These chemical-induced NPCs (ciNPCs) resemble mouse brain-derived NPCs regarding their proliferative and self-renewing abilities, gene expression profiles, and multipotency for different neuroectodermal lineages in vitro and in vivo. Further experiments reveal that alternative cocktails with inhibitors of histone deacetylation, glycogen synthase kinase, and TGF-ß pathways show similar efficacies for ciNPC induction. Moreover, ciNPCs can also be induced from mouse tail-tip fibroblasts and human urinary cells with the same chemical cocktail VCR. Thus our study demonstrates that lineage-specific conversion of somatic cells to NPCs could be achieved by chemical cocktails without introducing exogenous factors.