Rozkalne Anete, Adkin Carl, Meng Jinhong, Lapan Ariya, Morgan Jennifer E, Gussoni Emanuela
1 Program in Genomics and Division of Genetics, Boston Children's Hospital , Boston, MA 02115.
Hum Gene Ther. 2014 Jan;25(1):73-81. doi: 10.1089/hum.2013.126. Epub 2013 Dec 19.
Abstract Stem cell transplantation is being tested as a potential therapy for a number of diseases. Stem cells isolated directly from tissue specimens or generated via reprogramming of differentiated cells require rigorous testing for both safety and efficacy in preclinical models. The availability of mice with immune-deficient background that carry additional mutations in specific genes facilitates testing the efficacy of cell transplantation in disease models. The muscular dystrophies are a heterogeneous group of disorders, of which Duchenne muscular dystrophy is the most severe and common type. Cell-based therapy for muscular dystrophy has been under investigation for several decades, with a wide selection of cell types being studied, including tissue-specific stem cells and reprogrammed stem cells. Several immune-deficient mouse models of muscular dystrophy have been generated, in which human cells obtained from various sources are injected to assess their preclinical potential. After transplantation, the presence of engrafted human cells is detected via immunofluorescence staining, using antibodies that recognize human, but not mouse, proteins. Here we show that one antibody specific to human spectrin, which is commonly used to evaluate the efficacy of transplanted human cells in mouse muscle, detects myofibers in muscles of NOD/Rag1(null)mdx(5cv), NOD/LtSz-scid IL2Rγ(null) mice, or mdx nude mice, irrespective of whether they were injected with human cells. These "reactive" clusters are regenerating myofibers, which are normally present in dystrophic tissue and the spectrin antibody is likely recognizing utrophin, which contains spectrin-like repeats. Therefore, caution should be used in interpreting data based on detection of single human-specific proteins, and evaluation of human stem cell engraftment should be performed using multiple human-specific labeling strategies.
摘要 干细胞移植正在作为多种疾病的潜在治疗方法进行测试。直接从组织标本中分离或通过分化细胞重编程产生的干细胞，在临床前模型中需要对安全性和有效性进行严格测试。具有免疫缺陷背景且在特定基因中携带额外突变的小鼠的可用性，有助于在疾病模型中测试细胞移植的有效性。肌营养不良症是一组异质性疾病，其中杜氏肌营养不良症是最严重和最常见的类型。基于细胞的肌营养不良症治疗已经研究了几十年，正在研究的细胞类型有很多，包括组织特异性干细胞和重编程干细胞。已经建立了几种肌营养不良症的免疫缺陷小鼠模型，其中注射从各种来源获得的人类细胞以评估其临床前潜力。移植后，通过免疫荧光染色检测植入的人类细胞的存在，使用识别人类而非小鼠蛋白质的抗体。在这里，我们表明，一种通常用于评估移植的人类细胞在小鼠肌肉中功效的抗人血影蛋白特异性抗体，可检测到NOD/Rag1(null)mdx(5cv)、NOD/LtSz-scid IL2Rγ(null)小鼠或mdx裸鼠肌肉中的肌纤维，无论它们是否注射了人类细胞。这些“反应性”簇是正在再生的肌纤维，它们通常存在于营养不良组织中，血影蛋白抗体可能识别含有血影蛋白样重复序列的抗肌萎缩蛋白。因此，在基于检测单一人类特异性蛋白质来解释数据时应谨慎，并且应使用多种人类特异性标记策略来评估人类干细胞的植入情况。
