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（与之前提交的更改标记为 阴影文本 ） 项目 1 将测试质子放射治疗以超快剂量率（> 60 Gy/sec），称为 FLASH-PRT，与标准剂量率（<1 戈瑞/秒）质子放射治疗（S-PRT），同时使正常肠道组织免受急性和迟发毒性的影响。 成功完成拟议的研究将有助于更好地理解差异的机制 F-PRT 与 S-PRT 在分子、遗传和有机体水平上的影响相比较，并将定义 在初步发表的研究中，我们证明了启动临床试验所需的参数。 与 S-PRT 相比，F-PRT 提高了模型小鼠模型的总体存活率，并改善了晚期阶段 毒性，主要是纤维化，同时，F-PRT 在控制纤维化方面与 S-PRT 等效。 使用单细胞 RNA 测序 (scRNAseq) 的研究揭示了同种异体移植同基因肿瘤的生长。 上皮干细胞/祖细胞对 F-PRT 反应中基因表达谱的有趣差异 与 S-PRT 相比，这同时降低了对祖细胞增殖的抑制作用。 目标 1，我们将定义剂量测定和生物物理参数，以最大程度地保护正常组织 使用同源侧翼和原位模型以及基因工程研究 F-PRT 的抗肿瘤作用 PanCa 的小鼠模型（GEMM）然后我们将链接执行。 使用这些优化的剂量递增研究 与 S-PRT 相比，F-PRT 中确定的参数和局部 RT 可提高总体生存率 .在目标 2 中，我们 将描绘正常肠和肝组织（包括上皮）的差异反应机制， 血管、免疫和 循环 将细胞转为 S-PRT 和 F-PRT，使用 scRNAseq 对差异进行解卷积 在目标 3 中，我们将使用具有组织特异性的 GEMM。 删除 p53（上皮细胞与内皮细胞）以研究上皮细胞和内皮细胞的作用 分别针对 S-PRT 和 F-PRT 对急性和晚期毒性的反应以及上皮细胞减少 屏障损失。 我们还将测试 Wnt/β-catenin 和 R-Spondin 信号传导在介导 F- PRT 保留正常肠上皮 该项目将受益于概念性，并为此做出贡献。 从 scRNA-seq 收集的信息将为皮肤毒性实验提供信息。 和项目 2 中的祖细胞命运，包括上皮特异性和肉瘤的犬试验结果。 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 项目 3 将产生碳和质子辐照的肠道和 PanCa 样本。 将通过项目 1 进行分析。最后，项目 4 将开发一种笔形束扫描方法，我们将使用该方法 用于目标 1 下的剂量递增实验。