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.

抽象的淋巴管肌瘤病（LAM）是与硬化性硬化有关的疾病。两者都是由于 TSC1或TSC2基因中的遗传或零星突变，其作用为阴性 MTOR路径的调节器。不受控制的MTORC1活性导致肿瘤肺部异常平滑肌细胞（LAM细胞）的增殖呼吸，复发性气胸以及肺组织结构和功能的丧失原发性在女性中。 LAM的第一个也是唯一的FDA批准治疗方法是免疫抑制剂 Sirolimus，自2015年以来由Pfizer销售。它是当前的护理标准，并通过抑制 mtorc1。西罗莫司（Sirolimus 响应者，由于其免疫抑制特性和妊娠而导致的严重不良事件 C类，限制其在育龄女性中的使用。那是很高的未满足的医疗需求为LAM和TS开发替代和安全的治疗选择。我们已经确定了治疗 MiRNA302B模仿LAM/TS的潜在新疗法。使用鼠肺损伤模型，我们的合作者Hao Shen能够证明使用MirNA302B进行了非目标治疗中性脂质乳液的模仿改善了肺功能，宿主恢复和肺泡上皮再生小鼠。我们还展示了人类中的“停滞” AT2-AT1转变状态林，表明AT2适应性受损可能导致肺泡结构的丧失。我们的目标这笔赠款是靶向miRNA302B模拟脂质的肺上皮的投资通过增强AT2细胞再生来治疗LAM的纳米颗粒（LNP）效率。目标1 由miRNA-302B模拟脂质纳米颗粒的体外表征和调查体内几种靶向策略。我们的PI Jake Brenner博士将设计和在他的生物工程实验室中构建纳米颗粒。我们将执行3D器官实验在LNP [miR302b]存在下的人和小鼠AT2细胞随后进行了体内研究比较不同的靶向策略，以使用转基因到达所需的组织和细胞类型 LAM模型，由我们的其他PI教授Vera Krymskaya开发。使用最有效的定位策略，AIM 2将是使用相同的转基因鼠模型的体内概念验证研究怀孕前后，回答我们的治疗是否会阻止领空的问题扩大并导致肺恢复。气管内运动后，我们将观察小鼠4和8周 LNP [miR302b]给药。终点将包括肺功能，BALF分析，QPCR和为疾病相关标记和细胞类型染色的组织学肺切片。所有合作者是各自领域的专家，因此能够成功完成该项目，汇集了许多独特而创新的方面，以治疗这种毁灭性的罕见疾病。