Project 1: FLASH vs. Standard radiotherapy for treatment of PDAC and sparing normal intestine tissues

项目 1：FLASH 与标准放疗治疗 PDAC 并保护正常肠道组织

• 批准号: 10333798
• 负责人: Constantinos Koumenis
• 金额: $ 48.11万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-15 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10333798
• 关键词: Acute; Apoptosis; Blood Cells; Blood Vessels; Canis familiaris; Carbon; Cell Proliferation; Cells; Characteristics; Clinical Management; Clinical Trials; Columnar Cell; Coupled; Data; Development; Diagnosis; Disease; Dose; Dose-Rate; Endothelial Cells; Endothelium; Epithelial; Epithelial Cells; Exhibits; Fibrosis; Fractionation; Gene Expression; Gene Expression Profile; Genetic Determinism; Genetically Engineered Mouse; Growth; Immune; Immune response; Immunotherapy; Inflammatory Response; Injury; Interferon Type II; Intestines; Knock-out; Knockout Mice; LGR5 gene; Link; Liver; Lung; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Modality; Modeling; Molecular; Molecular Genetics; Mus; Normal tissue morphology; Organ; Organism; Pancreatic Adenocarcinoma; Play; Protons; Publishing; Radiation Tolerance; Radiation therapy; Resectable; Role; Sampling; Scanning; Scheme; Signal Transduction; Skin; Small Intestines; Survival Rate; TP53 gene; Testing; Text; Therapeutic; Time; Tissues; Toxic effect; Unresectable; antitumor effect; base; beta catenin; biophysical properties; cadherin 5; carcinogenesis; experimental study; gastrointestinal; gastrointestinal epithelium; improved; intestinal crypt; intestinal epithelium; mouse model; novel; pancreatic neoplasm; particle; progenitor; proton therapy; response; sarcoma; senescence; single-cell RNA sequencing; stem; stem cell population; stem cell proliferation; stem cells; translational study; tumor; tumor growth; tumorigenesis; villin

SUMMARY (changes from previous submission denoted with shaded text ) Project 1 will test the overall hypothesis that Proton Radiotherapy delivered in ultra-fast dose rates (>60 Gy/sec), termed FLASH-PRT, controls Pancreatic tumor growth equally well as Standard dose rate (<1 Gy/sec) Proton Radiotherapy (S-PRT), while sparing normal intestinal tissue from acute and delayed toxicity. Successful completion of the proposed studies will lead to better mechanistic understanding of the differential effects of F-PRT compared to S-PRT at the molecular, genetic and organismic level and will define the parameters necessary for the initiation of clinical trials. In preliminary published studies, we demonstrated that compared to S-PRT, F-PRT increases overall survival of model mouse models and also ameliorates late stage toxicity, primarily fibrosis. At the same time, F-PRT was shown to be equipotent to S-PRT in controlling the growth of allografted syngeneic tumors. Studies using single-cell RNA- sequencing (scRNAseq), reveal intriguing differences in gene expression profiles in the response of epithelial stem/progenitor cells to F-PRT compared to S-PRT which coincide with a reduction in the inhibitory effect on progenitor cell proliferation. In Aim 1, we will define the dosimetric and biophysical parameters which deliver maximum normal tissue sparing and anti-tumor effect of F-PRT using syngeneic flank and orthotopic models and Genetically Engineered Mouse model (GEMM) of PanCa. We will then link perform a dose-escalation study using these optimized parameters and focal RT to determine in F-PRT improves overall survival compared to S-PRT . In Aim 2, we will delineate the mechanism of differential response of normal intestinal and liver tissues including epithelium, vascular, immune and circulating cells to S-PRT and F-PRT, using scRNAseq to deconvolute the differential patterns of gene expression elicited by the two modalities. In Aim 3, we will use GEMM with tissue-specific deletion of p53 (epithelium vs. endothelium) to investigate the role of epithelial and endothelial cells respectively, in the response to S-PRT and F-PRT on acute and late toxicity coupled with reduced epithelial barrier loss. We will also test the involvement of the Wnt/β-catenin and R-Spondin signaling in mediating F- PRT sparing of normal intestinal epithelium . This project will benefit from, and contribute to, conceptual advances in the other projects. Information garnered from scRNA-seq will inform experiments on skin toxicity and progenitor cell fate in Project 2, including the canine trial on sarcoma. Results from epithelial-specific and endothelial-specific p53 knockout mice in this project will guide experiments in Projects 2 and Project 3 in sarcoma and lung. Project 3 will generate Carbon and Proton-irradiated intestinal and PanCa samples which will be analyzed by Project 1. Finally, Project 4 will develop a pencil-beam scanning approach which we will employ in the dose-escalation experiments under Aim 1.

摘要（从前提交表示的更改 阴影文字 ） 项目1将测试以超快速剂量率递送的质子放射疗法的总体假设（> 60 gy/sec）称为Flash-prt，控制胰腺肿瘤的生长与标准剂量率同样良好（<1 Gy/sec）质子放射疗法（S-PRT），同时从急性和延迟毒性中保留正常的肠道组织。 成功完成拟议的研究将导致对差异的更好理解 在分子，遗传和有机水平上与S-PRT相比，F-PRT的影响，将定义 临床试验倡议所需的参数。在初步发表的研究中，我们证明了 与S-PRT相比，F-PRT增加了模型小鼠模型的总体存活率，并改善后期 毒性，原发性纤维化。同时，F-PRT被证明与S-PRT相当 同种异体合成肿瘤的生长。使用单细胞RNA测序（SCRNASEQ）的研究揭示了 上皮茎/祖细胞对F-PRT的反应中基因表达谱的差异 与S-PRT相吻合，该S-prt与抑制性对祖细胞增殖的影响的降低相吻合。在 AIM 1，我们将定义剂量学和生物物理参数，这些参数提供最大的正常组织 F-PRT使用同步侧面和原位模型以及基因工程的抗肿瘤效应 panca的鼠标模型（GEMM）。然后我们将链接执行 使用这些优化的剂量提升研究 与S-prt相比 。在AIM 2中，我们 将描述正常肠道和肝组织差异反应的机制，包括上皮， 血管，免疫和 循环 细胞到S-PRT和F-PRT，使用scrnaseq反应差分 两种模态引起的基因表达模式。在AIM 3中，我们将使用GEMM与组织特异性 缺失p53（epitherium v​​s. Hothotherium）研究上皮细胞和内皮细胞的作用 在对急性和晚期毒性的对S-PRT和F-PRT的响应中，加上上皮降低 障碍损失。 我们还将测试Wnt/β-catenin和R-Spondin信号传导的参与 普通肠上皮的prt弹药 。该项目将从概念上受益，并为 其他项目的进步。从scrna-seq获得的信息将告知有关皮肤毒性的实验 项目2中的祖细胞命运，包括肉瘤的犬试验。来自上皮特异性的结果 该项目中的内皮特异性P53敲除小鼠将指导项目2和项目3的实验 肉瘤和肺。项目3将产生碳和质子辐射的肠和panca样品 将通过项目1分析。最后，项目4将开发一种铅笔梁扫描方法 在AIM 1下采用剂量降低实验。