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

项目总结/摘要 H3 K27 M突变型弥漫性中线胶质瘤（DMG）是一种毁灭性的小儿脑肿瘤。手术切除很困难 因为这些肿瘤位于脆弱的中脑/脑干位置，并渗透到关键结构。尽管 在数十年的临床试验中，H3 K27 M突变型DMG被认为是一致致命的，中位总生存期为 9-11个月放射治疗本身对发育中的儿童大脑有很高的毒性， 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MALT 1对H3 K27 M突变型DMG发病机制的影响。MALT 1是 CARMA/CARD-BCL 10-MALT 1（CBM）信号体的效应分子，一种细胞质蛋白复合物， 驱动下游促存活NF-κ B转录活性。MALT 1具有支架和蛋白酶 活动我们的实验室和其他实验室已经证明，MALT 1蛋白酶活性促进癌细胞增殖， 在多种癌症类型中的存活力、增殖和迁移/侵袭。除了在癌细胞中的作用外， MALT 1也是免疫细胞反应的关键调节因子，我们的实验室正在研究MALT 1对免疫细胞反应的影响。 高级别胶质瘤肿瘤微环境（TME）中的MALT 1蛋白酶。根据初步数据，我 假设抑制MALT 1蛋白水解活性可以在H3 K27 M突变体DMG中提供双重益处， 通过抑制癌细胞内MALT 1的直接作用和通过抑制MALT 1蛋白酶的作用 在TME的免疫细胞内，以促进抗肿瘤免疫激活。 在本提案的目的1中，我们将评估MALT 1蛋白酶活性对H3 K27 M突变体DMG的影响 TME。用于该目的的实验技术将包括使用鼠模型、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MALT 1蛋白酶的影响 H3 K27 M突变型DMG细胞的恶性特征的抑制，使用一系列体外分析。最后，在Aim 3，我们将在我们的原位/同基因组中分析MALT 1蛋白酶抑制的影响，有或没有辐射， H3 K27 M-突变体DMG模型。 总体而言，该项目评估了MALT 1蛋白酶的抑制作为H3 K27 M的潜在治疗方法。 突变体DMG完成这份提案将为我提供一个杰出的研究培训， 科学家和医生科学家团队。这次培训经验将为我成为一名医生做好准备- 研究儿科癌症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.