Elucidation of Tumor Resistance Mechanisms in Tuberous Sclerosis Complex-Associated Renal Angiomyolipoma for the Design of Novel Nanotherapies

阐明结节性硬化症相关肾血管平滑肌脂肪瘤的肿瘤抵抗机制，用于设计新型纳米疗法

• 批准号: 10585048
• 负责人: Dario Lemos
• 金额: $ 39.79万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10585048
• 关键词: Acceleration; Address; Adverse effects; Aftercare; Alleles; Anatomy; Angiomyolipoma; Apoptosis; Apoptotic; BCL-2 Protein; BCL1 Oncogene; BCL2 gene; BCL2L1 gene; BH3 Domain; BIRC3 gene; Benign; Binding Proteins; Biological Assay; Biology; Blood Vessels; CASP3 gene; CCI-779; CDKN1A gene; CRISPR/Cas technology; Cell Death; Cell Death Induction; Cell Survival; Cells; Cellular Morphology; Cessation of life; Characteristics; Combined Modality Therapy; Common Neoplasm; Complex; Cytoplasm; Cytostatics; Data; Development; Drug Combinations; Drug Delivery Systems; FRAP1 gene; Family; Generations; Genes; Genetic Engineering; Goals; Growth; Growth Factor; Half-Life; Hemorrhage; Human; Immunologic Deficiency Syndromes; In Vitro; Insulin; Interruption; Kidney; Kidney Failure; Kidney Neoplasms; Knock-out; Life; Mitochondria; Modeling; Molecular; Molecular Biology; Mutation; Organoids; PMAIP1 gene; Patients; Play; Population; Proteins; Rattus; Renal Angiomyolipoma; Reporting; Repression; Resistance; Resistance development; Retroperitoneal Hemorrhage; Rodent; Role; Rupture; SDZ RAD; Sampling; Signal Pathway; Smooth Muscle; Specificity; TSC1 gene; TSC2 gene; Testing; Time; Tissue Engineering; Tissues; Transcriptional Activation; Transgenic Mice; Transplantation; Tuberous Sclerosis; Tumor Promotion; Up-Regulation; Vascularization; Xenograft procedure; alpha Actin; antitumor effect; capsule; design; drug testing; efficacious treatment; efficacy evaluation; experimental study; glycoprotein NMB; in vivo; in vivo evaluation; induced pluripotent stem cell; inhibitor therapy; insight; kinase inhibitor; mTOR Inhibitor; melanocyte; mimetics; multicatalytic endopeptidase complex; nanocarrier; nanoparticle; nanotherapy; novel; novel therapeutics; pharmacologic; pre-clinical; premature; prevent; protein protein interaction; resistance mechanism; side effect; small hairpin RNA; tool; transcriptome; tumor

Summary A majority of patients with Tuberous Sclerosis Complex (TSC) develop benign kidney tumors known as renal angiomyolipoma (AML) that can cause renal insufficiency and spontaneous life-threatening hemorrhages. The main therapy for AML is everolimus, a rapamycin analog inhibitor of the kinase mTOR with cytostatic activity that only partially reduces tumor size. AMLs become stable over time, and tumor re-growth is often occurs after treatment is interrupted due to side effects. Therefore, there is an urgent need to elucidate mechanisms of tumor resistance for the development of more efficacious therapies. Efforts to recapitulate AML experimentally have failed for the past 20+ years, precluding the study of AML biology. To address this problem, we have used genetically engineered patient-derived induced pluripotent stem cells (iPSCs) to generate AML organoids. Organoids generated from iPSCs carrying biallelic inactivating mutations in the TSC2 locus (i.e. TSC2-/-) faithfully recapitulated key anatomical and molecular features of human kidney AML (reported in Hernandez JOR et al. Nat Commun. 2021 Nov 11;12(1):6496). Some of those features included the presence of myomelanocytic AML-like cells co-expressing smooth muscle and melanocyte markers, and the transcriptional activation of signaling pathways shared with kidney AML. Transplantation of TSC2-/- AML organoids into the kidneys of immunodeficient rodents resulted in fully vascularized human AML xenografts for mechanistic studies and for drug testing testing in vivo. Using these novel tools we identified potential mechanisms of tumor resistance driven by p21CIP1 and by BCL-2 apoptosis modulators, preventing AML cell death induced by rapalogs. Our in vivo experiments also indicated that drug delivery via nanocarriers may increase the efficacy of anti-tumor therapy while reducing undesired effects in other tissues. The objective of this proposal is to elucidate anti-apoptotic mechanisms driven by p21CIP1 and BCL-2 proteins for the development of novel anti-tumor therapies combining BCL-2 protein inhibitor drugs and rapalogs, that can be co-delivered using tissue-targeting nanoparticles. Our long-term goal is to design new therapies for AML with increased efficacy and specificity. The central hypothesis is that antiapoptotic mechanisms driven by p21CIP1 and BCL-2 apoptosis inhibitors sustain AML cell survival promoting tumor resistance to rapalog therapy. Our three aims are: Aim 1: To investigate anti-apoptotic mechanisms of tumor resistance driven by p21CIP1 in renal AML; Aim 2: To elucidate the role of IGFBP2 in stabilizing p21CIP1 promoting AML cell survival; Aim 3: To study the contribution of BCL-2 proteins to AML cell survival through pharmacologic blockage of BH3 domain interaction. Collectively, these studies will provide much needed insight into the mechanisms of AML and will assess the efficacy of BCL-2 inhibitor therapy alone or in combination with rapalogs in AML-targeting nanoparticles.

概括 大多数结节性硬化症复合物（TSC）患者出现良性肾脏肿瘤，称为肾脏肿瘤 血管肌脂肪瘤（AML）可能导致肾功能不全和自发威胁生命的出血。这 AML的主要疗法是依维莫司（Everolimus），这是一种具有细胞抑制酶MTOR的雷帕霉素抑制剂，具有细胞抑制活性 仅部分减少肿瘤的大小。随着时间的流逝，AML变得稳定，并且经常在 由于副作用，治疗被中断。因此，迫切需要阐明 肿瘤耐药性，以开发更有效的疗法。 在过去的20多年中，实验中概括AML的努力失败了，排除了AML的研究 生物学。为了解决这个问题，我们使用了基因设计的患者衍生的诱导多能茎 细胞（IPSC）生成AML器官。携带双重灭活的IPSC产生的类器官 TSC2基因座（即TSC2 - / - ）的突变忠实地概括了人类的关键解剖学和分子特征 肾脏AML（在Hernandez Jor等人的NatCommun。202111月11日； 12（1）：6496）。其中一些功能 包括存在共表达平滑肌和黑素细胞标记物的肌红细胞状AML样细胞， 以及与肾脏AML共享的信号通路的转录激活。 TSC2 - / - AML的移植 在免疫缺陷啮齿动物的肾脏中的器官导致完全血管化的人AML异种移植物 机械研究和用于体内的药物测试测试。使用这些新颖的工具，我们确定了潜力 由P21CIP1和BCL-2凋亡调节剂驱动的肿瘤耐药性机制，防止AML细胞 由Rapalogs诱发的死亡。我们的体内实验还表明，通过纳米载体输送药物可能 提高抗肿瘤疗法的功效，同时减少其他组织中不希望的影响。这个目的 建议是阐明由P21CIP1和BCL-2蛋白驱动的抗凋亡机制，以开发 结合Bcl-2蛋白抑制剂药物和Rapalogs的新型抗肿瘤疗法，可以使用 靶向组织的纳米颗粒。我们的长期目标是为AML设计新疗法，并提高功效 和特异性。中心假设是由P21CIP1和BCL-2驱动的抗凋亡机制 凋亡抑制剂维持AML细胞的存活，促进抗乳腺癌治疗的肿瘤性。我们的三个 目的是：目标1：研究由肾脏AML中P21CIP1驱动的肿瘤耐药性的抗凋亡机制； 目标2：阐明IGFBP2在稳定P21CIP1促进AML细胞存活中的作用；目标3：研究 Bcl-2蛋白通过BH3结构域相互作用的药理阻塞对AML细胞存活的贡献。 总的来说，这些研究将为AML机制提供急需的洞察力，并将评估 Bcl-2抑制剂疗法的功效或与AML靶向纳米颗粒中的Rapalog结合使用。