Bench to Beside and Back translational immuno-onocology-Cures

Bench to Beside and Back 转化免疫肿瘤学-Cures

• 批准号: 10926526
• 负责人: James L. Gulley
• 金额: $ 12.48万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10926526
• 关键词: Adolescent; Adult; Affect; Alternative Splicing; Antibody-drug conjugates; Antigens; Award; B-Cell Acute Lymphoblastic Leukemia; B-Lymphocytes; Back; Bone Tissue; Brain Neoplasms; CCR; CD19 gene; CD22 gene; Cancer Patient; Cancer cell line; Cell Line; Childhood; Childhood Leukemia; Clinical Trials; Data; Encyclopedias; Evaluation; Exons; Experimental Models; Extramural Activities; FDA approved; Goals; Hematologic Neoplasms; Human; Immune; Immune Targeting; Immunooncology; Immunotherapeutic agent; Immunotherapy; Laboratories; Libraries; Lymphoid Cell; Malignant Childhood Neoplasm; Malignant Neoplasms; Mediating; Messenger RNA; Methodology; NK cell therapy; Natural Killer Cells; Neuroblastoma; Neuroendocrine Tumors; Neurosecretory Systems; Open Reading Frames; Operative Surgical Procedures; PAX5 gene; Patient-Focused Outcomes; Patients; Pediatric Hospitals; Population Heterogeneity; Pre-Clinical Model; Procedures; Process; RNA Splicing; RNA-Binding Proteins; Recurrence; Refractory Disease; Relapse; Research; Resistance; Resistance development; Role; Sampling; Site; Soft tissue sarcoma; Solid Neoplasm; Source; T-Lymphocyte; Testing; Transcript; Transforming Growth Factor beta; Translating; Translational Research; Tumor Immunity; Untranslated RNA; Variant; Work; chemotherapy; childhood sarcoma; chimeric antigen receptor T cells; early phase clinical trial; effective therapy; experimental study; first-in-human; high throughput analysis; imprint; improved; improved outcome; manufacture; negative affect; novel; novel therapeutic intervention; participant enrollment; patient prognosis; phase I trial; prevent; protein expression; response; transcription factor; tumor; tumor microenvironment; vaccine development; vector; young adult

Four CCR projects are currently supported by this Cancer Moonshot Bench to Beside and Back award: 1. TME informed NK cell therapy for pediatric sarcomas, led by Rosandra Kaplan (CCR) and Timothy Cripe (Nationwide Children's Hospital). The overall goal of this project is to develop natural killer (NK) cell therapy with TGF-beta imprinting that can be augmented through manipulation of the immune suppressive tumor microenvironment (TME) leading to improved NK cell-mediated anti-tumor immunity, and serves as a potential effective therapy for patients with relapsed pediatric sarcoma. Bone and soft tissue sarcomas are one of the most common solid tumors in pediatric, adolescent/young adult (AYA) patients. Despite improved outcomes due to advances in surgical local control and multi-agent chemotherapy, prognosis for patients who develop recurrent, metastatic, or refractory disease remains poor. For these patients, new therapeutic approaches are urgently needed. The team at Nationwide Children's Hospital (NCH) has developed a genetically modified feeder cell line that enables extensive ex vivo propagation of highly-active human NK cells from various sources, and this methodology has been evaluated in early-phase clinical trials for hematologic malignancies and brain tumors. 2. Targeting neuroendocrine tumors with DLK1 directed immunotherapy, led by Nitin Roper (CCR) and John Maris (CHOP). During the first year of the BtB award we have focused on Aim 1 of our proposal: defining the anti-tumor activity of ADCT-701, a novel antibody drug conjugate targeting DLK1, in neuroendocrine neoplams. In particular, the goal of Aim 1 was to assess whether there is activity of ADCT-701 beyond neuroblastoma, which has been the focus of Dr. Maris' laboratory, our extramural collaborator. The results of our experiments in Year 1 demonstrate evidence of ADCT-701 anti-tumor activity in multiple ACC pre-clinical models. Thus, based on these data, ACC is an indication in our upcoming clinical trial: "A First in Human Phase I Trial with ADCT-701 in Neuroendocrine Neoplasms". The IND for ADCT-701 has been FDA approved and we expect to enroll patients in this clinical trial later this year. 3. B2B and Back: CD22 and CD19/22 CAR immunotherapies for childhood leukemia, led by Naomi Taylor (CCR) and Crystal Mackall (Stanford). The Taylor and Mackall teams are carrying out clinical trials with identical CD22 and CD19/CD22-bivalent CAR vectors but the two sites are using different manufacturing procedures to generate the CAR-transduced T cells that are being infused into patients. The current proposal is to test the hypothesis that variability in CAR-T cell immunotherapeutic potential is affected by variations in manufacturing platforms and can be predicted through evaluation of antigen-dependent CAR-T cell activity. High throughput analyses of these parameters will be leveraged to optimize CAR manufacturing, evaluate divergent patient outcomes, and enhance durable responses in pediatric leukemia patients. 4. Alternative splicing of CD22 following Inotuzumab, led by Nirali Shah (CCR) and Andrei Thomas-Tikhonenko (CHOP). Recent work from the Thomas-Tikhonenko lab has shown that CD22 protein expression is highly dependent on the inclusion of exon 2, which is where the open reading frame begins (Zheng et al., 2022). When exon 2 inclusion is negatively affected by aberrant splicing, the number of transcripts capable of being translated is significantly decreased, resulting in the emergence of CD22-low variants. To identify suitable experimental models for mechanistic studies, we quantitated total expression and reads spanning different exon 1 junctions in a panel of lymphoid cell lines from the Cancer Cell Line Encyclopedia (Ghandi et al., 2019). We observed that widely used NALM6 and REH cell lines differ in the relative abundance of the exon 2-skipping/non-coding CD22 mRNA variants, which were prominent in the former but not the latter. Using sgRNAs from the well-validated pooled Brunello library (Sanson et al. 2018) in NALM6 and REH cell lines we identified dozens of positive and negative regulators of CD22 expression that passed our stringent false discovery rate (FDR) cut-offs, with the CD22 gene predictably being the most significant regulator of its own protein expression. Further up on the list are known B cell-specific transcription factors PAX5 and EBF1, but also several RNA-binding proteins with potential roles in splicing. We are currently in the process of cross-referencing them with top correlates of CD22 exon 2 inclusion in B-ALL samples from the NCI. However, we are also prioritizing targets based on their known involvement in mRNA splicing as well as potential druggability.

目前，四个 CCR 项目得到了癌症登月台奖的支持： 1. TME 介绍了儿科肉瘤的 NK 细胞疗法，由 Rosandra Kaplan (CCR) 和 Timothy Cripe（全国儿童医院）领导。该项目的总体目标是开发具有 TGF-β 印记的自然杀伤 (NK) 细胞疗法，该疗法可以通过操纵免疫抑制性肿瘤微环境 (TME) 来增强，从而改善 NK 细胞介导的抗肿瘤免疫，并服务作为复发性小儿肉瘤患者的潜在有效疗法。骨和软组织肉瘤是儿科、青少年/年轻成人 (AYA) 患者中最常见的实体瘤之一。尽管由于手术局部控制和多药化疗的进步而改善了预后，但复发性、转移性或难治性疾病患者的预后仍然较差。对于这些患者来说，迫切需要新的治疗方法。全国儿童医院 (NCH) 的团队开发了一种转基因饲养细胞系，可实现各种来源的高活性人类 NK 细胞的广泛离体繁殖，并且该方法已在血液恶性肿瘤和癌症的早期临床试验中得到评估。脑肿瘤。 2. Nitin Roper (CCR) 和 John Maris (CHOP) 领导的 DLK1 定向免疫疗法针对神经内分泌肿瘤。在 BtB 奖的第一年，我们重点关注提案的目标 1：确定 ADCT-701（一种针对 DLK1 的新型抗体药物偶联物）在神经内分泌肿瘤中的抗肿瘤活性。特别是，目标 1 的目标是评估 ADCT-701 是否具有神经母细胞瘤以外的活性，这一直是我们的校外合作者 Maris 博士实验室的重点。我们第一年的实验结果证明了 ADCT-701 在多个 ACC 临床前模型中的抗肿瘤活性。因此，基于这些数据，ACC 是我们即将进行的临床试验的一个适应症：“首次使用 ADCT-701 治疗神经内分泌肿瘤的人体 I 期试验”。 ADCT-701 的 IND 已获得 FDA 批准，我们预计将在今年晚些时候招募患者参加这项临床试验。 3. B2B 和 Back：针对儿童白血病的 CD22 和 CD19/22 CAR 免疫疗法，由 Naomi Taylor (CCR) 和 Crystal Mackall（斯坦福大学）领导。 Taylor 和 Mackall 团队正在使用相同的 CD22 和 CD19/CD22 二价 CAR 载体进行临床试验，但这两个中心使用不同的制造程序来生成 CAR 转导的 T 细胞，并将其输注到患者体内。目前的提议是测试以下假设：CAR-T 细胞免疫治疗潜力的变异性受到制造平台变异的影响，并且可以通过评估抗原依赖性 CAR-T 细胞活性来预测。这些参数的高通量分析将用于优化 CAR 制造、评估不同的患者结果并增强儿童白血病患者的持久反应。 4. Inotuzumab 后的 CD22 选择性剪接，由 Nirali Shah (CCR) 和 Andrei Thomas-Tikhonenko (CHOP) 领导。 Thomas-Tikhonenko 实验室最近的工作表明，CD22 蛋白表达高度依赖于外显子 2 的包含，这是开放阅读框开始的地方 (Zheng et al., 2022)。当外显子 2 包含受到异常剪接的负面影响时，能够翻译的转录本数量显着减少，导致 CD22-low 变体的出现。为了确定适合机制研究的实验模型，我们对来自癌细胞系百科全书的一组淋巴细胞系中跨越不同外显子 1 连接的总表达和读数进行了定量（Ghandi 等人，2019）。我们观察到广泛使用的 NALM6 和 REH 细胞系在外显子 2 跳跃/非编码 CD22 mRNA 变体的相对丰度方面存在差异，这在前者中很突出，但在后者中则不然。在 NALM6 和 REH 细胞系中，使用经过充分验证的 Brunello 文库（Sanson 等人，2018）中的 sgRNA，我们鉴定了数十种 CD22 表达的正负调节因子，这些调节因子通过了我们严格的错误发现率 (FDR) 截止值，其中可以预见，CD22 基因是其自身蛋白质表达最重要的调节因子。名单中更靠前的是已知的 B 细胞特异性转录因子 PAX5 和 EBF1，还有几种在剪接中具有潜在作用的 RNA 结合蛋白。我们目前正在将它们与 NCI 的 B-ALL 样本中包含的 CD22 外显子 2 的最高相关性进行交叉引用。然而，我们还根据已知的 mRNA 剪接参与以及潜在的成药性来优先考虑目标。