Bench to Beside and Back translational immuno-onocology-Cures

目前有四个CCR项目得到了“癌症登月计划”奖项的支持：TME报告了由Rosandra Kaplan （CCR）和Timothy Cripe （national Children's Hospital）领导的针对儿童肉瘤的NK细胞疗法。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NK细胞介导的抗肿瘤免疫，并作为复发性儿童肉瘤患者的潜在有效治疗方法。骨和软组织肉瘤是儿童、青少年/青年（AYA）患者中最常见的实体肿瘤之一。尽管手术局部控制和多药化疗的进展改善了预后，但复发、转移或难治性疾病患者的预后仍然很差。对于这些患者，迫切需要新的治疗方法。全国儿童医院（NCH）的研究小组已经开发出一种转基因饲养细胞系，能够从各种来源广泛地体外繁殖高活性的人类NK细胞，这种方法已经在血液恶性肿瘤和脑肿瘤的早期临床试验中得到了评估。2. 由Nitin Roper （CCR）和John Maris （CHOP）领导的DLK1定向免疫治疗靶向神经内分泌肿瘤。该项目的一个中心目标是发现和优先考虑儿童癌症的免疫治疗靶点。由于DLK1蛋白在许多儿童和成人神经内分泌（NE）肿瘤中高表达，但正常表达受限，因此表面受体δ样非规范缺口配体1 （DLK1）已被优先考虑作为靶标。基于一种新型dlk1定向抗体-药物偶联物（ADCT-701）治疗神经母细胞瘤的临床前数据，研究小组建议对成人和儿童NE肿瘤患者进行研究，以进一步开发dlk1定向免疫疗法。DLK1在许多儿童和成人NE肿瘤中高表达的发现具有重要意义，因为神经内分泌肿瘤难以治疗，需要新的治疗方法。DLK1是神经母细胞瘤未分化状态的关键介质，神经母细胞瘤在DLK1基因缺失的情况下最终分化。提出的工作有可能导致加深对重要疾病过程的理解，并导致新的治疗干预。3. B2B和Back：儿童白血病的CD22和CD19/22 CAR免疫疗法，由Naomi Taylor （CCR）和Crystal Mackall （Stanford）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，但这两个地点使用不同的制造程序来产生CAR转导的T细胞，并将其注入患者体内。目前的建议是验证CAR-T细胞免疫治疗潜力的变异性受到制造平台变化的影响，并且可以通过评估抗原依赖性CAR-T细胞活性来预测这一假设。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，并增强儿科白血病患者的持久反应。4. 由Nirali Shah （CCR）和Andrei Thomas-Tikhonenko （CHOP）领导的Inotuzumab后CD22的选择性剪接。本项目旨在探索B-ALL肿瘤细胞中CD22 mRNA剪接改变（删除外显子2）导致CD22蛋白表达缺失的机制，从而导致这些肿瘤细胞对CD22特异性免疫疗法产生获得性耐药性。该项目结合了基于CD22的临床试验的临床样品的特征，以及CD22剪接调节的体外和体内研究；总之，这些研究可以确定新的治疗策略来强迫B-ALL细胞中CD22蛋白的表达，并改善抗CD22免疫治疗。

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 suppres sive 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, prog nosis 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 immunother apy, led by Nitin Roper (CCR) and John Maris (CHOP). A central goal of this project is to discover and prioritize immunotherapeutic targets in childhood cancers. Surface receptor Delta Like Non-Canonical Notch Ligand 1 (DLK1) has been prioritized as a target due to the pro tein's high expression in many pediatric and adult neuroendocrine (NE) tumors but with restricted normal expression. Based on pre-clinical data demonstrating efficacy of a novel DLK1-directed antibody-drug conjugate (ADCT-701) in neuroblastoma, the investigator team pro poses to study adult and pediatric patients with NE tumors to further develop DLK1-directed immunotherapy. The discovery that DLK1 is hiqhly expressed in many pediatric and adult NE tumors is of high significance since neuroendocrine tumors are difficult to treat and novel therapeutics are needed. DLK1 is a key mediator of the undifferentiated state in neuroblastoma, and neuroblastomas terminally differenti ate upon genetic depletion of DLK1. The proposed work has the potential to result in enhanced understanding of an important disease process and lead to a new therapeutic intervention. 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 po tential is affected by variations in manufacturing platforms and can be predicted through evaluation of antigen-dependent CAR-T cell activ ity. 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). This project is designed to explore the mechanism(s) by which altered splicing of CD22 mRNA, to delete Exon 2, leads to loss of CD22 protein expression in B-ALL tumor cells, and thus leads to the acquired resistance of those tumor cells to CD22-specific immunotherapies. The project is an attractive blend of characterization of clinical samples from CD22-based clinical trials, and in vitro plus in vivo studies of the regulation of CD22 splicing; together, these studies could identify new therapeutic strategies to force CD22 protein expression in B-ALL cells and improve anti-CD22-immunotherapy.