Mesenchymal Stem Cell Therapy: a1 antitrypsin deficiency

间充质干细胞治疗：a1抗胰蛋白酶缺乏症

• 批准号: 6602918
• 负责人: CATHERINE M VERFAILLIE
• 金额: $ 36.4万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6602918
• 关键词: alpha 1 antitrypsin deficiency; bone marrow; cell differentiation; cell population study; cell proliferation; flow cytometry; laboratory mouse; liver cells; liver disorder; liver function; liver metabolism; mesenchyme; nonhuman therapy evaluation; pluripotent stem cells; stem cell transplantation; tissue /cell culture

DESCRIPTION (provided by applicant): Alpha-l-antitrypsin (alpha1AT) is a plasma protein, produced in the liver that inhibits elastase, alpha1AT-deficiency is an autosomal recessive disorder that affects approximately 105 individuals in the US. Liver injury in alpha1AT-deficiency is caused by the hepatotoxic effects of mutant alpha1AT retained within the endoplasmic reticulum (ER) of hepatocytes, and emphysema by uninhibited proteolytic damage to elastic tissue in the lung parenchyma. Orthoptic liver transplantation is the only curative therapy for alpha1AT-mediated liver disease. If a reliable source of hepatocytes were available, they could conceivably be used to replace 20-25% of the host liver, elevating plasma levels of alpha1AT to greater than 10 muM, and preventing pulmonary toxicity due to alpha1AT-deficiency. We have identified multipotent adult progenitor cells, or MAPC, that can be cultured from human, mouse and rat bone marrow (BM). Single MAPC differentiate into mesodermal, neuroectoderm-like cells and functioning hepatocyte-like cells in vitro. MAPC do not form tumors when infused IV, IM or SQ, but differentiate in response to local cues into hematopoietic cells and epithelium of lung, intestine and liver. For MAPC derived hepatocytes to be suitable for therapy of alpha1AT, or other liver disorders, better characterization of the in vitro differentiation process will be needed. In addition, we will need to demonstrate that MAPC-derived cells function like hepatocytes in vivo. We propose three aims to determine whether MAPC-derived hepatocytes may be a source of cells to treat alpha1AT-deficiency mediated pulmonary and/or liver disease. Studies in SA1 will further demonstrate that hepatocyte like cells can be generated from bone marrow derived MAPC, and develop methods to select progenitors/precursors for hepatocytes generated during culture. Selection of such progenitors will serve two purposes: characterization of differentiation from pluripotent adult stem cells to hepatocytes, and generation of a potential source of cells for effective transplantation in liver disease. Studies described in SA2 that will assess all functions of mature hepatocytes in spheroid cultures should help confirm that hepatocytes generated from BM MAPC are similar to hepatocytes derived from primary liver. Studies in SA3 will establish whether MAPC themselves, MAPC-derived hepatocyte progenitors or mature hepatocytes can restore liver function in vivo, in the setting of hepatocyte cell death and in the setting of abnormal liver function but without liver cell death. This should lay the groundwork for future studies testing whether MAPC or MAPC-derived hepatocyte progenitors/hepatocytes can serve as a suitable source of cells for therapy of alpha1AT, a single gene defect associated with lung damage and/or liver failure.

描述（由申请人提供）： α-L-抗胰蛋白酶 (α1AT) 是一种血浆蛋白，在肝脏中产生，可抑制弹性蛋白酶。α1AT 缺乏症是一种常染色体隐性遗传疾病，在美国影响大约 105 个人。 α1AT 缺乏症的肝损伤是由保留在肝细胞内质网 (ER) 内的突变型 α1AT 的肝毒性作用引起的，而肺气肿是由肺实质弹性组织不受抑制的蛋白水解损伤引起的。正交肝移植是α1AT介导的肝病的唯一治疗方法。如果有可靠的肝细胞来源，它们可以用来替代 20-25% 的宿主肝脏，将 alpha1AT 的血浆水平提高到 10 muM 以上，并预防由于 alpha1AT 缺乏而引起的肺毒性。我们已经鉴定出可以从人类、小鼠和大鼠骨髓 (BM) 中培养的多能成体祖细胞 (MAPC)。单个 MAPC 在体外分化为中胚层、神经外胚层样细胞和功能性肝细胞样细胞。当静脉注射、肌内注射或 SQ 输注时，MAPC 不会形成肿瘤，而是根据局部信号分化为造血细胞以及肺、肠和肝的上皮细胞。为了使 MAPC 衍生的肝细胞适合治疗 alpha1AT 或其他肝脏疾病，需要更好地表征体外分化过程。此外，我们需要证明 MAPC 衍生细胞在体内的功能与肝细胞相似。我们提出了三个目标，以确定 MAPC 衍生的肝细胞是否可以作为治疗 alpha1AT 缺陷介导的肺部和/或肝脏疾病的细胞来源。 SA1 的研究将进一步证明肝细胞样细胞可以从骨髓来源的 MAPC 中产生，并开发为培养过程中产生的肝细胞选择祖细胞/前体的方法。选择此类祖细胞将有两个目的：表征从多能成体干细胞向肝细胞的分化，以及产生用于肝病有效移植的潜在细胞来源。 SA2 中描述的研究将评估球状培养物中成熟肝细胞的所有功能，应有助于确认从 BM MAPC 生成的肝细胞与源自原代肝脏的肝细胞相似。 SA3 的研究将确定 MAPC 本身、MAPC 衍生的肝细胞祖细胞或成熟肝细胞是否可以在肝细胞死亡的情况下以及肝功能异常但没有肝细胞死亡的情况下恢复体内肝功能。这应该为未来的研究奠定基础，测试MAPC或MAPC衍生的肝细胞祖细胞/肝细胞是否可以作为治疗α1AT的合适细胞来源，α1AT是一种与肺损伤和/或肝衰竭相关的单基因缺陷。