Real-time monitoring of circulating pancreatic tumor cells and clusters

实时监测循环胰腺肿瘤细胞和簇

基本信息

批准号：
10219169
负责人：
Erica Carpenter
金额：
$ 15万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10219169
关键词：
Acoustics Address Antibodies Antigens Area Biological Assay Biological Markers Blood Blood Tests Blood donor Blood specimen Cancer Etiology Cell Count Cell Line Cell Separation Cell surface Cells Cessation of life Clinical Clinical Research Colorectal Neoplasms Development Diagnosis Diagnostic radiologic examination Disease Disease Progression Distal FDA approved Fluorescence-Activated Cell Sorting Goals Heterogeneity Human Image Industry Laboratories Leukocytes Libraries Malignant Neoplasms Malignant neoplasm of pancreas Mammary Neoplasms Measurement Measures Mediating Methods Microfluidics Modeling Molecular Molecular Analysis Molecular Profiling Monitor Mus Neoplasm Circulating Cells Neoplasm Metastasis Operative Surgical Procedures Pancreatic Ductal Adenocarcinoma Pancreatic Ductal Carcinoma Patient Monitoring Patients Phenotype Preparation Prostatic Neoplasms RNA Recovery Recurrence Sampling Sensitivity and Specificity Site Sorting - Cell Movement Specificity Stains Survival Rate Technology Testing Therapeutic Time Tracer Travel X-Ray Computed Tomography base biomarker discovery biomarker panel biomarker signature burden of illness care outcomes cohort improved industry partner liquid biopsy metastatic process mouse model novel novel strategies pancreatic cancer cells pancreatic cancer patients pancreatic ductal adenocarcinoma model pancreatic neoplasm pre-clinical prognostic programs real time monitoring research clinical testing specific biomarkers standard of care tool transcriptome sequencing treatment response tumor

项目摘要

Pancreatic ductal adenocarcinoma (PDA), which has an overall five-year survival rate of 6%, is predicted to become the second leading cause of cancer-related death in the US. 80% of PDA patients who have surgery will suffer a recurrence likely due to undetectable metastases present at diagnosis; yet standard of care radiographic imaging and blood-tests can be imprecise and are not predictive of metastatic burden. Circulating tumor cells (CTCs) are thought to mediate much of the metastatic process, and while FDA-approved tests are established for enumeration of CTCs shed into the blood by breast, prostate, and colorectal tumors, such tests lack sufficient sensitivity for pancreatic tumors. Recently, we showed that: PDA CTC heterogeneity promoted a more aggressive phenotype; CTC clusters were associated with higher metastatic potential; and seeding at distal sites required more than one clone during the course of metastatic disease progression. This proposal seeks to develop and optimize a liquid biopsy for the measurement of single and clustered CTCs and their molecular signatures, in order to address the urgent unmet need for non-invasive clinical monitoring of PDA patient metastatic burden, disease progression, and treatment response. This industry/academic partnership will bring together Becton Dickinson (BD)'s strengths in rare cell capture and molecular analysis, Johns Hopkins' PDA antigen discovery programs, and Penn's pre-clinical PDA models, robust clinical PDA programs, and dedicated Circulating Tumor Material Laboratory. First, technologies for efficient enrichment and sorting of CTC clusters with high purity, recovery and viability will be optimized so that high quality RNA can be isolated for downstream molecular analyses (RNA-seq). The BD Focus, which is a rare cell enrichment platform being developed at BD, will be used for pre-enrichment and sorting using a combination of cell surface antibody and RNA-based probes (i.e., BD's unique “nanoflare” technology). Second, a panel of candidate CTC biomarkers will be finalized and validated in PDA pre-clinical mouse models as well as patients with metastatic PDA. Finally, we will establish proof of concept for the CTC biomarkers to predict and monitor the metastatic burden of a cohort of PDA patients starting at diagnosis and continuing after commencement of therapy, and at time points coinciding with monitoring CT scans. Single cell and clustered CTC counts and molecular signature will be correlated with metastatic burden, and survival. The ability to enrich and perform molecular characterization of CTC single cells and clusters will provide a novel and crucial technology to detect and monitor metastasis in PDA patients so that the appropriate therapeutic strategies can be utilized in order to increase overall survival rates.

胰腺导管腺癌 (PDA) 的总体五年生存率为 6%，预计将成为美国 80% PDA 患者癌症相关死亡的第二大原因。接受手术的患者可能会因诊断时存在无法检测到的转移而遭受复发；放射照相成像和血液测试的护理标准可能不精确，并且不能预测循环肿瘤细胞（CTC）被认为介导了大部分转移过程，虽然 FDA 批准的测试用于计数通过乳房流入血液中的 CTC，前列腺肿瘤和结直肠肿瘤，此类测试对胰腺肿瘤缺乏足够的敏感性。结果表明：PDA CTC 异质性促进了更具攻击性的表型；与较高的转移潜力相关；并且在远端部位播种需要多个克隆。该提案旨在开发和优化转移性疾病进展的液体。用于测量单个和簇 CTC 及其分子特征的活检，以便解决 PDA 患者转移性无创临床监测的迫切未满足需求这种行业/学术合作将带来负担、疾病进展和治疗反应。结合 Becton Dickinson (BD) 在稀有细胞捕获和分子分析方面的优势，约翰·霍普金斯大学 PDA 抗原发现计划、Penn 的临床前 PDA 模型、强大的临床 PDA 计划以及专门的循环肿瘤材料实验室。第一，高效富集和分选技术。具有高纯度、高回收率和高活力的CTC簇将被优化，从而可以得到高质量的RNA 分离用于下游分子分析 (RNA-seq)，这是一种罕见的细胞富集。 BD 正在开发的平台将用于使用细胞组合进行预富集和分选表面抗体和基于 RNA 的探针（即 BD 独特的“nanoflare”技术）。候选 CTC 生物标志物将在 PDA 临床前小鼠模型以及最后，我们将为 CTC 生物标志物进行预测建立概念验证。并监测一组 PDA 患者从诊断开始到诊断后的转移负担治疗开始时，以及与监测单细胞和 CT 扫描一致的时间点。聚集的 CTC 计数和分子特征将与转移负担和生存相关。富集和执行 CTC 单细胞和簇分子表征的能力将提供一种新颖的方法以及检测和监测 PDA 患者转移的关键技术，以便采取适当的治疗可以利用策略来提高总体存活率。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Flow Cytometric Methods for Circulating Tumor Cell Isolation and Molecular Analysis.

用于循环肿瘤细胞分离和分子分析的流式细胞术方法。

DOI：
10.1007/978-3-319-55947-6_5
发表时间：
2024-09-13
期刊：
Advances in experimental medicine and biology
影响因子：
0
作者：
N. Bhagwat;E. Carpenter
通讯作者：
E. Carpenter

An integrated enrichment system to facilitate isolation and molecular characterization of single cancer cells from whole blood.

一种集成富集系统，有助于从全血中分离单个癌细胞并进行分子表征。

DOI：
发表时间：
2018
期刊：
影响因子：
0
作者：
Yu, Liping;Sa, Silin;Wang, Ling;Dulmage, Keely;Bhagwat, Neha;Yee, Stephanie S;Sen, Moen;Pletcher Jr, Charles H;Moore, Jonni S;Saksena, Suraj;Dixon, Eric P;Carpenter, Erica L
通讯作者：
Carpenter, Erica L

Enumeration, Dielectrophoretic Capture, and Molecular Analysis of Circulating Tumor Cells.

循环肿瘤细胞的计数、介电泳捕获和分子分析。

DOI：
发表时间：
2017
期刊：
影响因子：
0
作者：
Yee, Stephanie S;Carpenter, Erica L
通讯作者：
Carpenter, Erica L

Transcriptional profiling of single tumour cells from pleural effusions reveals heterogeneity of epithelial to mesenchymal transition and extra-cellular matrix marker expression.

胸腔积液中单个肿瘤细胞的转录谱揭示了上皮间质转化和细胞外基质标志物表达的异质性。

DOI：
发表时间：
2022-07
期刊：
影响因子：
0
作者：
Sen, Moen;Hausler, Ryan M;Dulmage, Keely;Black, Taylor A;Murphy, William;Pletcher Jr, Charles H;Wang, Ling;Chen, Chang;Yee, Stephanie S;Bornheimer, Scott J;Maxwell, Kara N;Stanger, Ben Z;Moore, Jonni S;Thompson, Jeffrey C;Carpenter, Erica L
通讯作者：
Carpenter, Erica L

A Multianalyte Panel Consisting of Extracellular Vesicle miRNAs and mRNAs, cfDNA, and CA19-9 Shows Utility for Diagnosis and Staging of Pancreatic Ductal Adenocarcinoma.

由细胞外囊泡 miRNA 和 mRNA、cfDNA 和 CA19-9 组成的多分析物组合显示出对胰腺导管腺癌的诊断和分期的实用性。

DOI：
发表时间：
2020
期刊：
影响因子：
0
作者：
Yang, Zijian;LaRiviere, Michael J;Ko, Jina;Till, Jacob E;Christensen, Theresa;Yee, Stephanie S;Black, Taylor A;Tien, Kyle;Lin, Andrew;Shen, Hanfei;Bhagwat, Neha;Herman, Daniel;Adallah, Andrew;O'Hara, Mark H;Vollmer, Charles M;Katona, Bryson
通讯作者：
Katona, Bryson