Advances in application of induced pluripotent stem cells in childhood genetic diseases / 中国生物制品学杂志

@#Somatic cell reprogramming has developed rapidly in the field of modern biology. Induced pluripotent stem cells(iPSCs)obtained through somatic cell reprogramming are not only capable of self-renewal,but also have multidirectional differentiation potential,which plays an important role in disease modeling and regenerative medicine. This paper reviewed the gene reprogramming technology,the disease models of iPSCs and the application prospects of iPSCs in childhood genetic diseases,so as to provide a reference for the application of iPSCs in the research of mechanism and treatment of various genetic diseases.

Homology-based repair induced by CRISPR-Cas nucleases in mammalian embryo genome editing

Recent advances in genome editing, especially CRISPR-Cas nucleases, have revolutionized both laboratory research and clinical therapeutics. CRISPR-Cas nucleases, together with the DNA damage repair pathway in cells, enable both genetic diversification by classical non-homologous end joining (c-NHEJ) and precise genome modification by homology-based repair (HBR). Genome editing in zygotes is a convenient way to edit the germline, paving the way for animal disease model generation, as well as human embryo genome editing therapy for some life-threatening and incurable diseases. HBR efficiency is highly dependent on the DNA donor that is utilized as a repair template. Here, we review recent progress in improving CRISPR-Cas nuclease-induced HBR in mammalian embryos by designing a suitable DNA donor. Moreover, we want to provide a guide for producing animal disease models and correcting genetic mutations through CRISPR-Cas nuclease-induced HBR in mammalian embryos. Finally, we discuss recent developments in precise genome-modification technology based on the CRISPR-Cas system.

Develop a 3D neurological disease model of human cortical glutamatergic neurons using micropillar-based scaffolds

Establishing an effective three-dimensional (3D) culture system to better model human neurological diseases is desirable, since the human brain is a 3D structure. Here, we demonstrated the development of a polydimethylsiloxane (PDMS) pillar-based 3D scaffold that mimicked the 3D microenvironment of the brain. We utilized this scaffold for the growth of human cortical glutamatergic neurons that were differentiated from human pluripotent stem cells. In comparison with the 2D culture, we demonstrated that the developed 3D culture promoted the maturation of human cortical glutamatergic neurons by showing significantly more MAP2 and less Ki67 expression. Based on this 3D culture system, we further developed an disease-like model of traumatic brain injury (TBI), which showed a robust increase of glutamate-release from the neurons, in response to mechanical impacts, recapitulating the critical pathology of TBI. The increased glutamate-release from our 3D culture model was attenuated by the treatment of neural protective drugs, memantine or nimodipine. The established 3D human neural culture system and TBI-like model may be used to facilitate mechanistic studies and drug screening for neurotrauma or other neurological diseases.

Research progress cellular reprogramming technology and induced pluripotent stem cells for retinal diseases treatment / 中华实验眼科杂志

Current advances in cellular reprogramming technology has demonstrated that the identity of a cell can be converted by the use of master transcription factors to reprogram the transcriptome. Notably,this allows us to convert somatic cells into induced pluripotent stem cells (iPSCs),providing a feasible method to generate patient-specific pluripotent stem cells. This technology was firstly discovered by Shinya Yamanaka's group in 2006. The initial iPSCs were formed by the induction of dedifferentiation in mouse fibroblasts using transcription factors:Oct4,Sox2, Klf4 and c-Myc. This approach has tremendous medical potentials to revolutionize the way we study and develop treatment for ocular diseases. Here we reviewed the potential of using patient-specific iPSCs for 3D disease modeling and various types of retinal disease modeling,cell replacement therapy and clinical trials,high-throughput screening test and drug toxicity testing. We also discussed the recent development of direct reprogramming and the future direction for utilising iPSCs and cellular reprogramming technology for eye research.

From Bench to Market: Preparing Human Pluripotent Stem Cells Derived Cardiomyocytes for Various Applications

Human cardiomyocytes (CMs) cease to proliferate and remain terminally differentiated thereafter, when humans reach the mid-20s. Thus, any damages sustained by myocardium tissue are irreversible, and they require medical interventions to regain functionality. To date, new surgical procedures and drugs have been developed, albeit with limited success, to treat various heart diseases including myocardial infarction. Hence, there is a pressing need to develop more effective treatment methods to address the increasing mortality rate of the heart diseases. Functional CMs are not only an important in vitro cellular tool to model various types of heart diseases for drug development, but they are also a promising therapeutic agent for cell therapy. However, the limited proliferative capacity entails difficulties in acquiring functional CMs in the scale that is required for pathological studies and cell therapy development. Stem cells, human pluripotent stem cells (hPSCs) in particular, have been considered as an unlimited cellular source for providing functional CMs for various applications. Notable progress has already been made: the first clinical trials of hPSCs derived CMs (hPSC-CMs) for treating myocardial infarction was approved in 2015, and their potential use in disease modeling and drug discovery is being fully explored. This concise review gives an account of current development of differentiation, purification and maturation techniques for hPSC-CMs, and their application in cell therapy development and pharmaceutical industries will be discussed with the latest experimental evidence.

CRISPR/Cas9-mediated targeted gene correction in amyotrophic lateral sclerosis patient iPSCs

Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease with cellular and molecular mechanisms yet to be fully described. Mutations in a number of genes including SOD1 and FUS are associated with familial ALS. Here we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts of familial ALS patients bearing SOD1 and FUS mutations, respectively. We further generated gene corrected ALS iPSCs using CRISPR/Cas9 system. Genome-wide RNA sequencing (RNA-seq) analysis of motor neurons derived from SOD1 and corrected iPSCs revealed 899 aberrant transcripts. Our work may shed light on discovery of early biomarkers and pathways dysregulated in ALS, as well as provide a basis for novel therapeutic strategies to treat ALS.

Aplicaciones méiicas de las células pluripotentes inducidas paciente-específicas / Medical applicaiions of induced pluripotent cells specific-patient / Aplicações méicaas das células pluripotentes induzidas paciente-específicas

Los recientes avances en la implementación de estrategias de reprogramación genética en células somáticas para la producción de células pluripotentes inducidas (iPS), abren la posibilidad de generar células pluripotentes para estudios del desarrollo embrionario y la diferenciación celular, herramientas para detección in vitro de nuevos medicamentos y evaluación de su eficacia y toxicidad, desarrollo de modelos in vitro de enfermedades humanas y uso en terapia celular. Las iPS, son células que muestran características fenotípicas y funcionales similares a las observadas en células madre embrionarias, sin los cuestionamientos éticos y legales de la manipulación de embriones. En particular, la generación de las células pluripotentes inducidas paciente-específicas ha permitido dilucidar los procesos fisiopatológicos de diversas enfermedades genéticas de etiología conocida y desconocida, así como plantean la posibilidad de realizar terapia celular autóloga y terapia génica basada en células para la regeneración tisular dependiendo de las necesidades individuales.

Recent advances in the implementation of strategies of genetic reprogramming somatic cells to produce induced pluripotent cells (iPS), open the possibility of generating pluripotent cells for studies of embryonic development and cell differentiation, tools for in vitro detection of new drugs and evaluation of their efficacy and toxicity, in order to develop in vitro models of human disease and use in cell therapy. iPS cells are showing phenotypic and functional characteristics similar to those seen in embryonic stem cells, without the ethical and legal questionings of the experimental manipulation of embryos. In particular, generation of patient-specific pluripotent stem cells elucidate the pathophysiological processes of various genetic diseases of known and unknown aetiology, and raises the possibility of autologous cell therapy and cell-based gene therapy for tissue regeneration depending individual needs.

Os recentes avanços na implementação de estratégias de reprogramação genética em células somáticas para a produção de células pluripotentes induzidas (iPS), abrem a possibilidade de gerar células pluripotentes para estudos do desenvolvimento embrionário e a diferenciação celular, ferramentas para detecção in vitro de novos medicamentos e avaliação da sua eficácia e toxicidade, desenvolvimento de modelos in vitro de doenças humanas e uso em terapia celular. As iPS, são células que mostram características fenotípicas e funcionais similares às observadas em células tronco embrionárias, sem os questionamentos éticos e legais da manipulação de embriões. Em particular, a geração das células pluripotentes induzidas paciente-específicas tem permitido elucidar os processos fisiopatológicos de diversas doenças genéticas de etiologia conhecida e desconhecida, assim como estabelecem a possibilidade de realizar terapia celular autóloga e terapia gênica baseada em células para a regeneração tecidual dependendo das necessidades individuais.

Induced pluripotent stem cells and personalized medicine: current progress and future perspectives / 대한해부학회지

Generation of induced pluripotent stem cells (iPSCs) has revolutionized the field of regenerative medicine by providing researchers with a unique tool to derive disease-specific stem cells for study. iPSCs can self-renew and can differentiate into many cell types, offering a potentially unlimited source of cells for targeted differentiation into somatic effector cells. Hence, iPSCs are likely to be invaluable for therapeutic applications and disease-related research. In this review, we summarize the recent progress of iPSC generation that has been made with an emphasis on both basic and clinical applications including disease modeling, drug toxicity screening/drug discovery and cell replacement therapy.