Clinically suitable approach for gene-mediated therapy of cirrhosis.

摘要 肝脏持续性损伤可引起慢性炎症和细胞外基质沉积失调 (ECM)导致纤维化瘢痕组织的积累并最终导致肝硬化。纤维化是正常的伤口 愈合反应，在过量，它可以进一步损伤组织和激活促纤维化细胞，导致积极的 反馈回路瘢痕组织不是静态的，ECM重塑是一个动态和受调节的过程， 有望对纤维化疾病进行靶向干预。此外，最近的观察表明， 在去除损伤源后，晚期肝病的“逆转”。在肝硬化，积极清除 可能需要瘢痕组织来停止正反馈循环，为健康的人创造空间。 肝细胞生长。 实现这一点的一个假设是引入ECM调节酶，如胶原酶 基质金属蛋白酶-8（MMP-8），从而改变促纤维化组分与抗纤维化组分的平衡， 打破导致进一步纤维化沉积的正反馈循环， 肝细胞扩增这种方法导致纤维化病变明显减少，肝胶原蛋白减少， 在大鼠肝硬化模型中，减轻了肝负荷，改善了大体肝脏形态学，并改善了肝硬化相关症状。 然而，高效的腺病毒载体被用于这些概念验证研究。潜在毒性 阻碍了腺病毒载体在人类肝脏转导中的应用。因此，将这些发现翻译成 人类研究将需要更适合临床的方法，同时保持有效的基因递送。这 本申请提出了腺相关病毒载体的设计、构建、制造和表征。 (AAV)表达MMP-8的载体来实现这一点，并将该项目推进到临床研究。 本提案中包含的合作团队在GMP-AAV制造方面具有丰富的经验， 纤维化疾病的临床前模型，并具有监管和临床开发所需的专业知识， 将结果转化为肝硬化患者的人类基因转移研究。成功完成后 该项目的第二阶段将包括GMP载体生产和临床前 提交IND以支持临床研究所需的药理学和毒理学研究。

Abstract Persistent injury to the liver can cause chronic inflammation and dysregulated deposition of extracellular matrix (ECM), leading to accumulation of fibrotic scar tissue and eventually cirrhosis. While fibrosis is a normal wound healing response, in excess, it can further injure tissue and activate pro-fibrotic cells, resulting in a positive feedback loop. Scar tissue is not static, ECM remodeling is a dynamic and regulated process that holds promise for targeted intervention of fibrotic disease. Furthermore, recent observations suggest the potential for the 'reversal' of advanced liver disease upon removal of the source of injury. In cirrhosis, aggressive removal of scar tissue would likely be needed to halt the positive feedback loop and create space for healthy hepatocyte growth. One hypothesis for achieving this is the introduction of ECM regulating enzymes, such as the collagenase Matrix Metalloprotienase-8 (MMP-8), thus shifting the balance of pro-fibrotic versus anti-fibrotic components, breaking the positive feedback loop that leads to further fibrotic deposits and creating space for healthy hepatocyte expansion. This approach led to marked decreases in fibrotic lesions, decreased hepatic collagen burden, improved gross liver morphology, and cirrhosis-associated symptoms in rat cirrhosis models. However, highly efficient adenoviral vectors were used for those proof-of-concept studies. Potential toxicity hinders the use of adenoviral vectors for liver transduction in humans. Thus, translation of these findings to human studies will require a more clinically suitable approach, while maintaining efficient gene delivery. This application proposes the design, construction, manufacture, and characterization of an Adeno-associated viral (AAV) vector expressing MMP-8 to accomplish this and advanced this project to clinical studies. The collaborative team included in this proposal has extensive experience with GMP-AAV manufacturing, preclinical models for fibrotic disease, and has the regulatory and clinical development expertise required to translate the results into a human gene transfer study in patients with cirrhosis. Upon successful completion of this proposal, the second phase of this project will include GMP vector production and pre-clinical pharmacology and toxicology studies necessary to submit an IND to support clinical studies.