Tumor targeted engineered stem cells for treatment of lung cancer

该提案旨在探索和测试一种新的靶向溶瘤病毒细胞疗法，该疗法使用 呼吸道合胞病毒感染人骨髓间充质干细胞 治疗肺癌的病毒(RSV)，这是癌症中的头号杀手 全世界。尽管在治疗方面取得了进展，结果也有所改善，但5年生存率 非小细胞肺癌(NSCLC)的发病率仍然非常低(15%或更低)，这构成了 约85%的肺癌患者。如此低的存活率与肺癌 强调对经典化疗和放射治疗的异质性和耐药性的发展 开发新的非姑息性治疗策略的必要性。尽管有溶瘤作用 病毒疗法已经成为传统化疗和放射疗法的潜在替代品， 它们在肺癌上的应用仍然有限。此外，无法将溶瘤病毒靶向肿瘤 总的来说，仍然是一个关键障碍，特别是对肺癌来说，仍然是一个主要的未得到满足的需求。 针对肺癌的靶向溶瘤病毒细胞疗法的概念的动机来自 来自两个主要的发展。首先是间充质干细胞，这是一种众所周知的具有固有肿瘤的细胞类型。 热带特性，被发现对RSV高度敏感，感染率为90%，提示 利用RSV感染的MSCs进行肿瘤靶向和肿瘤杀伤。这一潜力是 受RSV感染的MSCs上调吲哚胺-2，3-双加氧酶(IDO)的挑战， 抑制肿瘤免疫，从而有助于肿瘤的形成。然而，人们发现IDO- 缺陷的hMSCs仍然对RSV高度敏感，同时减轻了肿瘤抑制作用。 第二，非结构蛋白1(Ns1)缺陷(Δns1)RSV在肺内复制时病毒滴度很高 肿瘤细胞，与正常的WI-38二倍体肺细胞相比。这些发现加在一起导致了 假设∆NS1RSV感染的IDO缺陷的MSCs将保持肿瘤趋向性和 将有助于开发一种针对肺癌的有针对性的溶瘤病毒细胞疗法。这 假设将在以下三个具体目标中得到检验。好了！ 在目标#1中，计划开发和鉴定稳定的IDO缺陷的hMSC细胞系，使用 CRISPR法检测野生型和工程化(Δ)Ns1的抗肿瘤活性 呼吸道合胞病毒感染的骨髓间充质干细胞(RMSCs)在体外和活检组织来源的3D多细胞瘤样培养中的作用 类肿瘤体外实验。在目标2中，建议评估RMSCs在体内的抗癌潜力。 免疫活性同种异体肺癌原位移植模型的建立 调节抗肿瘤免疫。在Aim#3中，RMSC在调节细胞凋亡中的作用 将对病毒抗原进行调查。 总体而言，靶向RMSC疗法有望提供高度创新和有效的 肺癌的治疗方法。私人投资总监处于独特的位置，可以进行拟议的 研究开发肺癌的新治疗方法，该方法可能适用于其他 肿瘤疾病。所有必要的方法、试剂和协作都已到位 成功完成这一拟议计划，在肺癌领域具有最高的临床影响。

This proposal aims to explore and test a novel targeted oncolytic viro-cell therapy using the mesenchymal stem cells (MSCs) infected by harmless common cold virus, the respiratory syncytial virus (RSV) for treatment of lung cancers, which is the number one killer among cancers worldwide. Despite advances in the treatment and improvement in outcomes, the 5-year survival rates remain very poor (15% or less) for non–small cell lung cancers (NSCLC), which constitute ~85% of all lung cancer patients. Such poor survival rates combined with lung cancer heterogeneity and development of resistance to classical chemo- and radio-therapies, underscore the need for development of novel non-palliative therapeutic strategies. Though oncolytic virotherapies have emerged as a potential alternative to traditional chemo- and radio-therapies, their application to lung cancers remain limited. Also, inability to target oncolytic viruses to tumors in general remains a critical barrier and particularly for lung cancers remains a major unmet need. The motivation for the concept of targeted oncolytic viro-cell therapy against lung cancer comes from two major developments. First, MSCs, a cell-type generally known to have intrinsic tumor tropic properties, were found to be highly susceptible to RSV, with >90% infection rate, suggesting the use of RSV-infected MSCs for both tumor targeting and tumor killing. This potential was challenged by upregulation of indoleamine-2, 3-dioxygenase (IDO) by RSV-infected MSCs, which suppresses tumor immunity and thus aid in tumorigenesis. However, it was found that IDO- deficient hMSCs remain highly susceptible to RSV while alleviating the tumor suppressive effect. Second, nonstructural protein 1 (NS1) deficient (ΔNS1) RSV replicates with high viral titer in lung tumor cells, compared to the normal WI-38 diploid lung cells. Together these findings have led to the hypothesis that the ∆NS1 RSV-infected IDO-deficient MSCs will retain tumor tropism and would lend themselves to develop a targeted oncolytic viro-cell therapy against lung cancers. This hypothesis will be tested in the following three specific aims. ! In aim #1, it is planned to develop and characterize a stable IDO-deficient hMSC cell line using the CRISPR method and examine the anti-tumor activity potential of wild type and engineered (ΔNS1) RSV-infected MSCs (RMSCs) in 3D multicell tumoroid cultures in vitro and biopsy-derived tumoroids ex vivo. In aim #2, it is proposed to assess the anti-cancer potential of RMSCs in vivo in immunocompetent syngeneic orthotopic model of lung cancer and evaluate their potential to modulate anti-tumor immunity. In aim #3 the effects of RMSC in modulating apoptosis induced by viral antigens will be investigated. Overall, the targeted RMSC therapy is expected to provide a highly innovative and effective approach for treatment of lung cancers. The PIs are uniquely positioned to conduct the proposed studies to develop a novel therapeutic approach for lung cancers, which may apply to other neoplastic diseases. All the required methods, reagents and collaborations are in place for the successful completion of this proposed program with highest clinical impact in the lung cancer field.