Design of a micro-RNA attenuated armed oHSV for the treatment of glioblastoma

脑恶性肿瘤的最具侵袭性的形式，多形性胶质母细胞瘤（GBM）代表特别急性的恶性肿瘤。 未满足的医疗需求GBM是高度侵袭性的，并且由于它们的免疫抑制而不响应免疫检查点抑制。 抑制性微环境经工程改造以选择性复制和杀死恶性GBM的溶瘤病毒 细胞和表达刺激抗肿瘤免疫的转基因是改善患者免疫功能的有吸引力的方法。 作为单一疗法和与抗PD-1治疗剂组合的结果。获得的初步数据 原型溶瘤性单纯疱疹病毒（oHSV）装备ULBP 3，ULBP 3是KLRK 1刺激受体的配体 在CD 8和NK细胞中表达，并具有溶瘤效力、微RNA衰减 由Oncorus设计的有效载荷容量在单次瘤内注射后显示出显著的生存益处， 在设计用于概括主要遗传改变的遗传诱导型GBM模型中注射， GBM的病理生物学此外，在一个肿瘤块中注射表达ULBP 3的oHSV导致了远隔 未注射肿瘤块中的抗肿瘤反应。这一结果是令人惊讶的，因为人类特异性ULBP 3 基因产物不识别小鼠KLRK 1受体，主要通过募集 巨噬细胞和小胶质细胞，表明ULBP 3在小鼠和人类的这些细胞上的新受体。 初步研究确定了组织因子，一种跨膜受体，在凝血和信号传导中起作用， GBM作为这种新的受体。我们的目的是研究ULBP 3在GBM模型中的作用模式，并证实这一点。 新颖的互动。我们建议进一步利用这些初步研究结果的设计优化武装oHSV 用于治疗GBM。作为第一步，我们将定制Oncorus oHSV的独特安全功能， 在健康脑组织中高度表达的micro-RNA，在GBM中丢失以插入互补的micro-RNA HSV复制所必需的多个基因中的靶序列，从而抑制病毒复制至恶性 细胞显示出强差异表达的microRNA 124、128和137可能是候选者。此外，本发明的目的是， 有效载荷，包括IL-12、PD-1和CD 47拮抗剂，已在GBM临床前模型中得到验证， 并且可以补充ULBP 3活性的蛋白将在GBM细胞系和肿瘤细胞中逐步测试， 在同基因GBM模型中和在最先进的可诱导GBM模型中的溶瘤活性、表达和体内功效 基因模型根据这些数据，有效载荷的优化组合将被引入我们的oHSV中， 为未来的人体试验创造最有效的载体和临床候选物。 !

The most aggressive form of brain malignancies, glioblastoma multiforme (GBM) represents a particularly acute unmet medical need. GBM are highly invasive and do not respond to immune checkpoint inhibition due to their suppressive microenvironment. Oncolytic viruses engineered to selectively replicate and kill malignant GBM cells and to express transgenes that stimulate antitumor immunity are attractive approaches to improve patient outcomes as monotherapy and in combination with anti-PD-1 therapeutics. Preliminary data achieved with a prototypical oncolytic herpes simplex virus (oHSV) armed with ULBP3, a ligand for KLRK1 a stimulatory receptor expressed in CD8 and NK cells and featuring the improvements in oncolytic potency, micro-RNA attenuation and payload capacity designed by Oncorus demonstrated a significant survival benefit after a single intratumoral injection in a genetically inducible model of GBM designed to recapitulate the major genetic alterations and pathobiology of GBM. Moreover, injection of oHSV expressing ULBP3 in one tumor mass resulted in an abscopal anti-tumor response in the uninjected tumor mass. This result was surprising since the human-specific ULBP3 gene product does not recognize the mouse KLRK1 receptor and primarily acts through the recruitment of macrophages and microglia, suggesting a novel receptor for ULBP3 on these cells both in mouse and human. Preliminary studies identified tissue factor, a transmembrane receptor with a role in coagulation and signaling in GBM as this new receptor. We aim to investigate the mode of action of ULBP3 in GBM models and confirm this novel interaction. We propose to further exploit these initial findings for the design of an optimized armed oHSV for the treatment of GBM. As a first step, we will tailor the unique safety features of Oncorus oHSV, the selection of micro-RNA highly expressed in healthy brain tissues and lost in GBM to insert complementary micro-RNA target sequences in multiple genes essential to HSV replication, thus restraining viral replication to malignant cells. MicroRNA 124, 128 and 137 that showed strong differential expression are likely candidates. Additionally, payloads, including IL-12, and PD-1 and CD47 antagonists, that have been validated in GBM preclinical models, and that may complement the activity of ULBP3 will be tested stepwise in GBM cell lines and tumor cells for oncolytic activity, expression and in vivo efficacy in syngeneic GBM models and in state-of-the-art inducible genetic models. From these data the optimized combination of payloads will be introduced into our oHSV in order to create the most potent vector and clinical candidate for future human trials. !