Department of Genetics, Faculty of Biology and Soil Sciences, Kazan Federal University, ul. Kremlevskaya 18, R-420008 Kazan, Russia.
Exp Biol Med (Maywood). 2011 Jan;236(1):91-8. doi: 10.1258/ebm.2010.010172. Epub 2010 Dec 16.
Current therapy of a number of neuropsychiatric maladies has only symptomatic modality. Effective treatment of these neuro-degenerative diseases, including amyotrophic lateral sclerosis (ALS), may benefit from combined gene/stem-cell approaches. In this report, mononuclear fraction of human umbilical cord blood cells (hUCBCs) were transfected by electroporation with dual plasmid constructs, simultaneously expressing vascular endothelial growth factor 165 (VEGF(165)) and human fibroblast growth factor 2 (FGF(2)) (pBud-VEGF-FGF(2)). These genetically modified hUCBCs were injected retro-orbitally into presymptomatic ALS transgenic animal models ((G)93(A) mice). Lumbar spinal cords of rodents were processed for immunofluoresent staining with antibodies against human nuclear antigen (HNA), oligodendrocyte-specific protein, S100, iba1, neuronal β(3)-tubulin and CD34. Co-localization of HNA and S100 was found in the spinal cord of mice after transplantation of genetically modified hUCBCs over-expressing VEGF-FGF(2). Double staining in control animals treated with unmodified hUCBCs, however, revealed HNA+ cells expressing iba1 and CD34. Neuron-specific β(3)-tubulin or oligodendrocyte-specific protein were not expressed in hUCBCs in either control or experimental mice. These results demonstrate that genetically naïve hUCBCs may differentiate into endothelial (CD34+) and microglial (iba1+) cells; however when over-expressing VEGF-FGF(2), hUCBCs transform into astrocytes (S100+). Autocrine regulation of VEGF and FGF(2) on hUCBCs, signal molecules from dying motor neurons in spinal cord, as well as self-differentiating potential may provide a unique microenvironment for the transformation of hUCBCs into astrocytes that eventually serve as a source of growth factors to enhance the survive potential of surrounding cells in the diseased regions.
目前，许多神经精神疾病的治疗方法只有对症疗法。通过基因/干细胞方法联合治疗这些神经退行性疾病，包括肌萎缩侧索硬化症（ALS），可能会有所受益。在本报告中，通过电穿孔将人脐血单核细胞（hUCBC）的单核细胞部分转染到同时表达血管内皮生长因子 165（VEGF（165））和人成纤维细胞生长因子 2（FGF（2））的双质粒构建体中（pBud-VEGF-FGF（2））。将这些基因修饰的 hUCBC 经眶后注射到有症状前的 ALS 转基因动物模型（（G）93（A）小鼠）中。用针对人核抗原（HNA）、少突胶质细胞特异性蛋白、S100、iba1、神经元β（3）-微管蛋白和 CD34 的抗体对啮齿动物的腰脊髓进行免疫荧光染色。在转染过表达 VEGF-FGF（2）的基因修饰 hUCBC 后，在小鼠的脊髓中发现了 HNA 和 S100 的共定位。然而，在用未经修饰的 hUCBC 治疗的对照动物中，双染色显示 HNA+细胞表达 iba1 和 CD34。在对照或实验小鼠的 hUCBC 中均未表达神经元特异性β（3）-微管蛋白或少突胶质细胞特异性蛋白。这些结果表明，未成熟的 hUCBC 可以分化为内皮细胞（CD34+）和小胶质细胞（iba1+）；然而，当过度表达 VEGF-FGF（2）时，hUCBC 转化为星形胶质细胞（S100+）。hUCBC 上的 VEGF 和 FGF（2）的自分泌调节、脊髓中死亡运动神经元的信号分子以及自我分化潜能可能为 hUCBC 转化为星形胶质细胞提供独特的微环境，最终成为生长因子的来源，以增强病变区域周围细胞的存活潜力。
