miRNA-Nanotechnology as a novel regenerative therapy for lymphangioleiomyomatosis

miRNA-纳米技术作为淋巴管平滑肌瘤病的新型再生疗法

基本信息

批准号：
10761353
负责人：
Jacob Brenner
金额：
$ 30.1万
依托单位：
BIOSPUTNIK LLC
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-25 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10761353
关键词：
3-Dimensional Acceleration Adverse effects Affect Age Alveolar Antibodies Biomedical Engineering CD47 gene Categories Cell Culture Techniques Cells Clinical Cyst DNA Data Disadvantaged Disease Disease Progression Drug Delivery Systems Emulsions Ensure Epithelial Cells Epithelium Equilibrium FDA approved FRAP1 gene Family Female Female of child bearing age Flow Cytometry Formulation Genetic Goals Grant Heterozygote Histologic Human Human Characteristics Immune response Immunosuppressive Agents Impairment In Vitro Inherited Intercept Intravenous Investigation Lesion Life Lipids Liposomes Lung Lung Lymphangioleiomyomatosis Lung diseases Lymphangioleiomyomatosis Marketing Measures Medical Mesenchyme MicroRNAs Microscopic Modeling Mus Mutation Nanotechnology Natural regeneration Organoids Pathway interactions Patients Persons Pharmaceutical Preparations Pneumothorax Pregnancy Prevention Proliferating Property Pulmonary function tests Qualifying RNA Rare Diseases Recovery Recurrence Research Route SDZ RAD Sampling Scientist Serious Adverse Event Shortness of Breath Signal Transduction Sirolimus Smooth Muscle Smooth Muscle Myocytes Stains Structure Surface System TSC1 gene TSC2 gene Testing Tissues Transgenic Mice Transgenic Model Transgenic Organisms Treatment Efficacy Tuberous Sclerosis Tumor Suppressor Genes United States Woman age related alveolar epithelium cell regeneration cell type clinical development cohort conditional knockout design design and construction epithelium regeneration experimental study fetal fitness gain of function improved in vivo innovation lipid nanoparticle lung development lung injury lung lesion microRNA delivery mouse model nanocarrier nanoparticle neoplastic novel novel therapeutics post pregnancy pregnant prepregnancy prevent pulmonary function pulmonary function decline rare genetic disorder regenerative therapy repaired restoration sex small molecule standard of care stem cells success targeted delivery targeted treatment tissue repair transdifferentiation

项目摘要

ABSTRACT Lymphangioleiomyomatosis (LAM) is a Tuberous sclerosis-related disorder. Both occur due to an inherited or sporadic mutation in either the TSC1 or TSC2 gene, which function as negative regulators of the mTOR pathway. Uncontrolled mTORC1 activity leads to the neoplastic proliferation of abnormal smooth muscle cells (LAM cells) in the lungs, progressive shortness of breath, recurrent pneumothoraxes, and loss of pulmonary tissue structure and function primarily in women. The first and only FDA-approved treatment for LAM is the immunosuppressant sirolimus, marketed since 2015 by Pfizer. It is the current standard-of-care and acts by inhibiting mTORC1. Sirolimus has several clinical disadvantages, including a considerable number of non- responders, severe adverse events due to its immunosuppressive properties and pregnancy category C, limiting its use in women of childbearing age. Thus, there is a high unmet medical need to develop alternative and safer treatment options for LAM and TS. We have identified treatment with miRNA302b mimics as a potential novel therapy for LAM/TS. Using a murine lung injury model, our collaborator Hao Shen was able to show that non-targeted treatment with miRNA302b mimics as neutral lipid emulsion improved lung function, host recovery, and alveolar epithelial regeneration mice. We also demonstrated a “stalled” AT2-AT1 transdifferentiation state in human LAM, suggesting that impaired AT2 fitness may contribute to loss of alveolar structure. Our goal for this grant is the investigation of pulmonary-epithelium targeted miRNA302b mimic lipid nanoparticles (LNP) efficacy for the treatment of LAM by enhancing AT2 cell regeneration. Aim 1 is composed of in vitro characterization of the miRNA-302b mimic lipid nanoparticle and investigation of several targeting strategies in vivo. Our PI Dr. Jake Brenner will design and construct the nanoparticles in his bioengineering lab. We will perform 3D organoid experiments with human and mouse AT2 cells in the presence of the LNP[miR302b] followed by an in vivo study comparing different targeting strategies to reach the desired tissue and cell type using a transgenic model of LAM, developed by our other PI Prof. Vera Krymskaya. Using the most effective targeting strategy, Aim 2 will be an in vivo proof-of-concept study using that same transgenic murine model pre and post-pregnancy to answer the question whether our treatment will prevent airspace enlargement and lead to lung recovery. We will observe mice for 4 and 8 weeks after intratracheal LNP[miR302b] administration. Endpoints will include lung function, BALF analysis, qPCR, and histological lung sections stained for the disease related markers and cell types. All collaborators are experts in their respective field and thus well equipped to successfully complete this project, bringing together a number of unique and innovative aspects to treat this devastating rare disease.

抽象的淋巴管平滑肌瘤病 (LAM) 是一种与结节性硬化症相关的疾病。 TSC1 或 TSC2 基因的遗传性或散发性突变，其功能为阴性 mTOR 通路的调节因子导致 mTORC1 活性失控。肺部异常平滑肌细胞（LAM 细胞）增殖，进行性呼吸困难呼吸、反复气胸以及肺组织结构和功能主要丧失 FDA 批准的第一个也是唯一一个针对女性的 LAM 治疗方法是免疫抑制剂。西罗莫司，自 2015 年起由辉瑞上市，是目前的治疗标准，通过抑制发挥作用。 mTORC1 有几个临床缺点，包括相当多的非- 反应者、由于其免疫抑制特性而导致的严重不良事件和怀孕 C 类，限制其在育龄妇女中的使用，因此，存在很高的未满足的医疗需求。为 LAM 和 TS 开发替代且更安全的治疗方案，我们已经确定了治疗方法。使用 miRNA302b 模拟小鼠肺损伤作为 LAM/TS 的潜在新疗法。模型中，我们的合作者Hao Shen 能够证明使用 miRNA302b 进行非靶向治疗模拟中性脂肪乳剂改善肺功能、宿主恢复和肺泡上皮我们还在人类体内展示了“停滞”的 AT2-AT1 转分化状态。 LAM，表明 AT2 适应性受损可能会导致肺泡结构丧失。这笔资助的目的是研究肺上皮靶向 miRNA302b 模拟脂质纳米粒子（LNP）通过增强 AT2 细胞再生来治疗 LAM 的功效。由 miRNA-302b 模拟脂质纳米颗粒的体外表征组成，我们的 PI 博士 Jake Brenner 将设计和研究几种体内靶向策略。我们将在他的生物工程实验室中构建纳米颗粒，进行 3D 类器官实验。在 LNP[miR302b] 存在的情况下与人和小鼠 AT2 细胞进行实验，然后进行体内研究使用转基因技术比较不同的靶向策略以达到所需的组织和细胞类型 LAM 模型，由我们的另一位 PI 教授 Vera Krymskaya 开发，使用最有效的靶向。策略，目标 2 将是使用相同转基因小鼠模型的体内概念验证研究怀孕前和怀孕后回答我们的治疗是否会阻止空域的问题我们将在气管内观察小鼠 4 周和 8 周。 LNP[miR302b] 管理终点包括肺功能、BALF 分析、qPCR 和对疾病相关标记物和细胞类型进行染色的组织学肺切片所有合作者。是各自领域的专家，因此有能力成功完成该项目，汇集了许多独特和创新的方面来治疗这种毁灭性的罕见疾病。