MALT1 protease as a regulator of anti-tumor immunity and tumor progression in diffuse midline glioma

项目摘要/摘要 H3K27M-突变型弥漫性中线胶质瘤(DMG)是一种毁灭性的儿童脑瘤。手术切除很困难 因为这些肿瘤位于微妙的中脑/脑干位置，并渗透到关键结构中。尽管 数十年的临床试验表明，H3K27M突变DMG被认为是一致致命的，总存活率的中位数为 9-11个月。放射治疗本身对发育中的儿童大脑有很大的毒性，它通过 2-3个月。因此，开发安全有效的DMG治疗方法至关重要。 我建议评估抑制MALT1在H3K27M突变DMG发病机制中的作用。Malt1是 CARMA/CARD-Bcl10-MALT1(CBM)信号小体的效应分子，是一种细胞质蛋白复合体， 驱动下游促进生存的核因子-kB转录活性。麦芽糖1同时具有支架和蛋白酶。 活动。我们的实验室和其他人已经证明了MALT1蛋白水解酶活性促进癌细胞 多种癌症类型的生存能力、增殖和迁移/侵袭。除了它在癌细胞中的作用外， MALT1也是免疫细胞反应的关键调节因子，我们的实验室正在调查 高级别胶质瘤微环境(TME)中MALT1蛋白的表达根据我的初步数据，我 假设抑制MALT1蛋白分解活性可以在H3K27M突变株DMG中提供双重好处，两者 通过在癌细胞内抑制MALT1的直接作用和通过抑制MALT1蛋白酶的作用 在免疫细胞内促进TME的抗肿瘤免疫激活。 在本提案的目标1中，我们将评估MALT1蛋白酶活性对H3K27M突变株DMG的影响 我也是。用于这一目的的实验技术将包括使用小鼠模型、单细胞RNA测序、 和多光谱免疫荧光。在本提案的目标2中，我们将评估MALT1蛋白酶的影响 用一系列体外分析方法抑制H3K27M突变DMG细胞的恶性特征。最后，在AIM中 3，我们将分析MALT1蛋白酶抑制在我们的原位/同基因移植中的影响，无论有没有辐射 H3K27M-突变型DMG模型。 总体而言，该项目评估了抑制MALT1蛋白酶作为H3K27M的一种潜在治疗方法。 突变的DMG。完成这项提议将为我提供杰出的研究培训 科学家和内科科学家组成的团队。这段培训经历将为我成为一名医生做好准备- 研究儿童癌症的分子基础以开发新的和改进的治疗方法的科学家。

PROJECT SUMMARY/ABSTRACT H3K27M-mutant diffuse midline glioma (DMG) is a devastating pediatric brain tumor. Surgical resection is difficult because these tumors are in delicate midbrain/brainstem locations and infiltrate into critical structures. Despite decades of clinical trials, H3K27M-mutant DMG is considered uniformly fatal, with a median overall survival of 9-11 months. Radiation therapy, which itself is highly toxic to the developing pediatric brain, extends survival by 2-3 months. Therefore, the development of safe and effective therapies for DMG is of utmost importance. I propose to evaluate the effect of inhibiting MALT1 on the pathogenesis of H3K27M-mutant DMG. MALT1 is the effector molecule of the CARMA/CARD-BCL10-MALT1 (CBM) signalosome, a cytoplasmic protein complex that drives downstream pro-survival NF-kB transcriptional activity. MALT1 possesses both scaffolding and protease activities. Our laboratory and others have demonstrated that MALT1 protease activity promotes cancer cell viability, proliferation, and migration/invasion in multiple cancer types. In addition to its role within cancer cells, MALT1 is also a critical regulator of immune cell responses, and our laboratory is investigating the impact of MALT1 protease in the tumor microenvironment (TME) of high-grade glioma. Based on my preliminary data, I hypothesize that inhibiting MALT1 proteolytic activity could provide dual benefit in H3K27M-mutant DMG, both via the direct effects of inhibiting MALT1 within the cancer cells and via the effects of inhibiting MALT1 protease within immune cells of the TME to promote anti-tumor immune activation. In Aim 1 of this proposal, we will evaluate the influence of MALT1 protease activity on the H3K27M-mutant DMG TME. Experimental techniques utilized in this aim will include use of murine models, single cell RNA sequencing, and multispectral immunofluorescence. In Aim 2 of this proposal, we will evaluate the impact of MALT1 protease inhibition on malignant features of H3K27M-mutant DMG cells, using a series of in vitro analyses. Finally, in Aim 3, we will analyze the impact of MALT1 protease inhibition, with or without radiation, in our orthotopic/syngeneic H3K27M-mutant DMG model. Overall, this project evaluates inhibition of MALT1 protease as a potential therapeutic approach for H3K27M- mutant DMG. Completion of this proposal will provide me with exceptional research training by an outstanding team of scientists and physician-scientists. This training experience will prepare me for a career as a physician- scientist who studies the molecular underpinnings of pediatric cancers to develop new and improved treatments.