Podvysotskaya V S, Grigor'eva E V, Malakhova A A, Minina J M, Vyatkin Y V, Khabarova E A, Rzaev J A, Medvedev S P, Kovalenko L V, Zakian S M
Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia Novosibirsk State University, Novosibirsk, Russia.
Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
Vavilovskii Zhurnal Genet Selektsii. 2025 Feb;29(1):15-25. doi: 10.18699/vjgb-25-03.
Parkinson's disease is a neurodegenerative disorder affecting dopaminergic neurons of the substantia nigra pars compacta. The known pathological genetic variants may explain the cause of only 5 % of cases of the disease. In our study, we found two patients with a clinical diagnosis of Parkinson's disease with the genetic variant c.1087G>T (p.Gly363Cys) of the LGR4 gene. The LGR4 gene encodes the membrane receptor LGR4 (leucine rich repeat containing G protein-coupled receptor 4) associated with the G protein. We hypothesize that the LGR4 gene may be either a direct cause or a risk factor for this disease, since it is one of the main participants of the WNT/β-catenin signalling pathway. This signalling pathway is necessary for the proliferation of neurons during their differentiation, which may lead to Parkinson's disease. To study the relationship between this genetic variant and Parkinson's disease, an ideal tool is a cellular model based on induced pluripotent stem cells (iPSCs) and their differentiated derivatives, dopaminergic neurons. We reprogrammed the peripheral blood mononuclear cells of the two patients with the c.1087G>T variant of the LGR4 gene with non-integrating episomal vectors expressing OCT4, SOX2, KLF4, LIN28, L-MYC and mp53DD proteins. The obtained seven lines of induced pluripotent stem cells were characterised in detail. The iPSCs lines obtained meet all the requirements of pluripotent cells, namely, they stably proliferate, form colonies with a morphology characteristic of human pluripotent cells, have a normal diploid karyotype, express endogenous alkaline phosphatase and pluripotency markers (OCT4, NANOG, SSEA-4 and SOX2) and are capable to differentiate into derivatives of the three germ layers. The iPSC lines obtained in this work can be used as a tool to generate a relevant model to study the effect of the pathological variant c.1087G>T of the LGR4 gene on dopaminergic neuron differentiation.
帕金森病是一种影响黑质致密部多巴胺能神经元的神经退行性疾病。已知的病理性基因变异仅能解释该疾病5%的病例病因。在我们的研究中，我们发现两名临床诊断为帕金森病的患者携带LGR4基因的c.1087G>T（p.Gly363Cys）基因变异。LGR4基因编码与G蛋白相关的膜受体LGR4（富含亮氨酸重复序列的G蛋白偶联受体4）。我们推测LGR4基因可能是该疾病的直接病因或危险因素，因为它是WNT/β-连环蛋白信号通路的主要参与者之一。该信号通路在神经元分化过程中的增殖是必需的，而这可能导致帕金森病。为了研究这种基因变异与帕金森病之间的关系，一个理想的工具是基于诱导多能干细胞（iPSC）及其分化衍生物多巴胺能神经元的细胞模型。我们用表达OCT4、SOX2、KLF4、LIN28、L-MYC和mp53DD蛋白的非整合附加型载体对两名携带LGR4基因c.1087G>T变异的患者的外周血单个核细胞进行重编程。对获得的七株诱导多能干细胞系进行了详细表征。所获得的iPSC系符合多能细胞的所有要求，即它们能稳定增殖，形成具有人类多能细胞特征形态的集落，具有正常的二倍体核型，表达内源性碱性磷酸酶和多能性标志物（OCT4、NANOG、SSEA-4和SOX2），并且能够分化为三个胚层的衍生物。本研究中获得的iPSC系可作为一种工具，用于生成相关模型，以研究LGR4基因的病理性变异c.1087G>T对多巴胺能神经元分化的影响。
