Novel Combination Therapy for Treatment and Prevention of PulmonaryLymphangioleiomyomatosis (LAM) and Tuberous Sclerosis Complex (TSC)

治疗和预防肺淋巴管平滑肌瘤病 (LAM) 和结节性硬化症 (TSC) 的新型联合疗法

• 批准号: 10697901
• 负责人: VERA P KRYMSKAYA
• 金额: $ 30万
• 依托单位: BIOSPUTNIK LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10697901
• 关键词: Acceleration; Adverse event; Affect; Age; Animals; Antibodies; Benign; Biological Products; Brain; Bronchoalveolar Lavage; C57BL/6 Mouse; Cancer Model; Categories; Cell Count; Cell Culture Techniques; Cell Line; Cells; Clinic; Clinical; Code; Combined Modality Therapy; Complex; Control Groups; Cyst; Cytoplasm; Data; Development; Disadvantaged; Disease; Dose; Ensure; Environment; Eye; FDA approved; FRAP1 gene; Female of child bearing age; Fibroblasts; Flow Cytometry; Genetic; Genetic Diseases; Goals; Grant; Growth; Heart; Human; Immune; Immune Evasion; Immunocompetent; Immunoglobulins; Immunohistochemistry; Immunologic Surveillance; Immunosuppression; Immunosuppressive Agents; Immunotherapy; In Vitro; Incubated; Inherited; Injections; Investigation; Kidney; Laboratories; Lesion; Libraries; Life; Literature; Lung; Lung Lymphangioleiomyomatosis; Lung Transplantation; Lung diseases; Lymphangioleiomyomatosis; Lymphatic Obstruction; Malignant Neoplasms; Medical; Mesenchymal; Modeling; Morphology; Mus; Mutation; Neoplasms; Nodule; Organ; PD-1 blockade; Pathway interactions; Patients; Pattern; Pennsylvania; Persons; Pharmaceutical Preparations; Pneumothorax; Pregnancy; Preparation; Prevention; Preventive treatment; Proliferating; Property; Publishing; Quantitative Reverse Transcriptase PCR; Rare Diseases; Recurrence; Regimen; Reporting; Research; Safety; Sequential Treatment; Shortness of Breath; Signal Transduction; Sirolimus; Skin; Smooth Muscle Myocytes; Structure; Surface; Suspensions; System; TSC1 gene; TSC1/2 gene; TSC2 gene; Tail; Testing; Therapeutic; Tissues; Translating; Transplant Recipients; Tuberous Sclerosis; Tumor Suppressor Genes; United States; Universities; Up-Regulation; Veins; Western Blotting; Woman; Work; anti-PD-1; anti-PD1 antibodies; anti-PD1 therapy; checkpoint inhibition; child bearing; clinical development; clinical translation; comparison group; curative treatments; efficacy evaluation; experimental study; gene function; immune checkpoint blockade; immunoregulation; improved; improved outcome; in vivo; intraperitoneal; lung lesion; mTOR Inhibitor; mouse model; mutant; neoplastic; novel; novel therapeutics; pembrolizumab; programmed cell death ligand 1; standard of care; time use; tumor

1 ABSTRACT 2 Tuberous sclerosis (TS), also called tuberous sclerosis complex, is a rare, multi-systemic genetic and often life- 3 threatening disease that causes benign tumors to grow in the brain and on other vital organs such as the kidneys, 4 heart, eyes, lungs, and skin. Lymphangioleiomyomatosis (LAM) is a TS-related tumor-like disorder. Both occur 5 as a consequence of an inherited or sporadic mutation in either the TSC1 or TSC2 gene, which function as 6 negative regulators of the mTOR pathway. Uncontrolled mTORC1 activity leads to the neoplastic proliferation of 7 abnormal smooth muscle cells (LAM cells) in the lungs, progressive shortness of breath, recurrent 8 pneumothoraxes, and loss of pulmonary tissue structure and function. In addition, published data suggests that 9 LAM cells may evade immune surveillance by upregulating the surface expression of programmed death-ligand 1 10 (PD-L1). LAM is a rare disease affecting women in childbearing age. The first and only FDA-approved treatment 11 for LAM is the immunosuppressant Rapamycin. It is the current standard-of-care and acts by inhibiting mTORC1. 12 Rapamycin has several clinical disadvantages, including a considerable number of non-responders, severe 13 adverse events due to its immunosuppressive properties and pregnancy category C, limiting its use in women of 14 childbearing age. Thus, there is a high unmet medical need to develop alternative and safer treatment options for 15 LAM and TS. We have identified Anti-PD1 checkpoint blockade as a potential novel therapy for LAM/TS. Using 16 our recently developed immunocompetent mouse model, the Co-PI Prof. Krymskaya was able to show that 17 treatment with an Anti-PD1 antibody significantly improved mouse surival. To advance immunetherapy for LAM, 18 our goal for this grant is the investigation of the mechanistic interplay between Rapamyin and Anti-PD1 antibodies 19 to harness the potential of a combination therapy in vitro and in vivo. Aim 1 will cover a set of in vitro studies, 20 investigating immune-relevant expression patterns and underlying cellular effects on mTOR and related signaling 21 pathways, since the relationship between the former and checkpoint blockade is not fully understood yet. We will 22 use immortalized TSC2-null 101, 102 and TSC2-expressing 103 cells before moving on to our expanded library 23 of primary human LAM-derived AML (LAMD) cells, originating from over 25 LAM patients. Investigating the effect 24 of Rapamycin and mTOR/Akt signalling will increase our understanding of the connection between those pathways 25 and PD-L1 upregulation, before moving on to Aim 2. The in vivo proof of concept will compare preventative and 26 therapeutic co-treatment with Anti-PD1 and Rapamying in an immunocompetent TSC2-null murine model 27 developed at the laboratory of Prof. Krymskaya at the University of Pennsylvania. We will evaluate prevention of 28 TSC2-null lesion growth, immunohistochemical and lung morphology changes and animal survival. This project 29 aims to develop a novel therapy for patients with the devastating diseases of LAM/TS. Based on Pembrolizumab’s 30 favorable safety profile in its approved indications, positive efficacy results in our studies would enable an 31 accelerated clinical development under FDA BPIC act for biologics and orphan disease designation for LAM/TS. 32

1篇摘要 2结节性硬化症（TS），也称为结节性硬化症，是一种罕见的，多系统遗传性，往往是终身的， 3威胁性疾病，导致良性肿瘤生长在大脑和其他重要器官，如肾脏， 4心脏、眼睛、肺和皮肤。淋巴管平滑肌瘤病（LAM）是一种与TS相关的肿瘤样疾病。均发生 5是由于TSC 1或TSC 2基因中的遗传性或散发性突变，其功能是 mTOR通路的6个负调节因子。不受控制的mTORC 1活性导致肿瘤性增殖， 7例肺部平滑肌细胞（LAM细胞）异常，进行性呼吸短促，复发 8例气胸，肺组织结构和功能丧失。此外，公布的数据表明， LAM细胞可能通过上调程序性死亡配体1的表面表达来逃避免疫监视。 10（PD-L1）。LAM是一种影响育龄女性的罕见疾病。第一个也是唯一一个FDA批准的治疗方法 11对于LAM是免疫抑制剂雷帕霉素。它是目前的标准治疗，通过抑制mTORC 1发挥作用。 12雷帕霉素有几个临床缺点，包括相当数量的无应答者，严重的 由于其免疫抑制特性和妊娠C类，13起不良事件，限制了其在以下女性中的使用： 14生育年龄因此，开发替代和更安全的治疗选择是高度未满足的医疗需求， 15、我和你我们已经确定抗PD 1检查点阻断作为LAM/TS的潜在新疗法。使用 16我们最近开发的免疫活性小鼠模型，Co-PI Krymskaya教授能够证明， 17用抗PD 1抗体治疗显著改善小鼠存活。为了推进LAM的免疫治疗， 我们的目标是研究Rapamyin和抗PD 1抗体之间的相互作用机制。 19利用体外和体内联合治疗的潜力。目标1将涵盖一系列体外研究， 20研究免疫相关的表达模式和对mTOR和相关信号传导的潜在细胞效应 21条途径，因为前者与检查站封锁之间的关系尚未完全了解。我们将 22使用无限增殖的TSC 2-null 101、102和表达TSC 2的103细胞，然后转移到我们的扩增文库 23个原代人LAM衍生的AML（LAMD）细胞，来源于超过25个LAM患者。调查影响 24雷帕霉素和mTOR/Akt信号传导将增加我们对这些途径之间联系的理解 25和PD-L1上调，然后转移到目标2。体内概念验证将比较预防性和 26在免疫活性TSC 2缺失鼠模型中用抗PD 1和Rapamying的治疗性共同治疗 27在宾夕法尼亚大学Krymskaya教授的实验室开发的。我们将评估预防 28 TSC 2-null病灶生长、免疫组织化学和肺形态学变化及动物存活率。这个项目 29旨在为患有LAM/TS的毁灭性疾病的患者开发一种新的疗法。基于Pembrolizumab的 在其批准的适应症中有30个有利的安全性特征，我们的研究中的积极疗效结果将使 31根据FDA BPIC法案加速LAM/TS生物制剂和孤儿病认定的临床开发。 32