Functional characterization of brain-colonizing breast cancer CTC subsets

脑定植乳腺癌 CTC 亚群的功能特征

• 批准号: 10249055
• 负责人: Dario Marchetti
• 金额: $ 32.68万
• 依托单位: UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10249055
• 关键词: Affect; Automobile Driving; Autopsy; Biological Assay; Biological Markers; Blood Volume; Blood specimen; Brain; Breast Cancer Patient; Breast Cancer Prevention; Breast Cancer Treatment; Breast Cancer cell line; Breast Cancer therapy; Cell Adhesion Molecules; Cell Proliferation; Cells; Clinic; Clinical; Competence; Complementary DNA; Country; Development; Diagnosis; Disease; Drug usage; ERBB2 gene; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Epithelial Cells; Flow Cytometry; Frequencies; Fright; Gene Expression; Gene Expression Profiling; Growth; Heparanase inhibitors; Homing; Human; In Vitro; Knowledge; Laboratories; Lentivirus; Link; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Metastatic breast cancer; Metastatic malignant neoplasm to brain; Molecular; Morbidity - disease rate; Neoplasm Circulating Cells; Neoplasm Metastasis; Neurofibromatoses; Neurofibromin 2; Oncogenic; PAK-1 kinase; Pathway interactions; Patients; Phenotype; Phosphorylation; Play; Positioning Attribute; Property; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Risk; Role; Seeds; Signal Transduction; Specimen; TACSTD1 gene; Technology; Testing; Therapeutic; Therapeutic Effect; Thermodynamics; Translational Research; Tumor Cell Line; Validation; Xenograft procedure; base; cancer subtypes; circulating biomarkers; cohort; combat; combinatorial; design; drug discovery; effective therapy; heparanase; high risk; improved; inter-institutional; malignant breast neoplasm; mortality; novel; novel therapeutics; pre-clinical; prevent; prognostic; prognostic assays; public health relevance; small hairpin RNA; small molecule; syndecan; targeted biomarker; targeted treatment; therapeutic target; tumor; tumorigenesis; tumorigenic; virtual

 DESCRIPTION (provided by applicant): Breast cancer brain metastasis (BCBM) is a devastating disease whose mechanisms are understudied and remain largely unknown. For example, while human epidermal growth factor receptor1 and 2 (EGFR and HER2) are high-risk predictors of BCBM, HER2/EGFR-targeted therapies have proven to be mostly ineffective against BCBM. Although notions that circulating tumor cells (CTCs) acting as "seeds" of intractable metastasis are established, virtually nothing is known about the properties and biomarkers of breast cancer brain-colonizing CTCs. Therefore, there is an urgent need for new CTC target biomarkers to combat BCBM. We have discovered CTCs that do not express the epithelial cell adhesion molecule (EpCAM-negative CTCs) and possess high BCBM competency in xenografts. These CTCs could not be detected by CellSearchTM, a FDA-cleared prognostic test which evaluates only EpCAM-positive CTCs. We found two promising markers in EpCAM-negative CTCs to predict BCBM in the clinic: Notch1, an important tumor-initiating cell biomarker, and heparanase (HPSE), a potent tumorigenic, angiogenic and pro-metastatic molecule. Our results indicate that BCBM-inducing CTCs have a "Brain Metastasis-Selected Marker (BMSM)" profile or HER2+/EGFR+/Notch1+/HPSE+ CTCs. Notch1 and HPSE can thus represent novel and specific CTC biomarkers. Furthermore, we detected highest expression of the tumor modulator Merlin (Neurofibromatosis type2) in Notch1/HPSE-expressing CTCs. The phosphorylation of Merlin (P-S518 Merlin) by PAK1 kinase resulted in oncogenic functions of Merlin which associate with BMSM CTC pathways and their regulation. Based on these findings, our hypothesis is that Notch1 and HPSE are novel CTC biomarkers that can predict the presence of primary breast cancer brain metastasis; and they can be potential therapeutic targets to prevent secondary BCBM. Objective of this proposal is to perform preclinical translational research and target validation to nominate HPSE and Notch1 as critical biomarkers of breast cancer brain-homing CTCs. We will study CTCs from HER2+ patients since this breast cancer subtype has a much higher than average risk of developing BCBM. Specifically, we will: 1) determine effects of therapeutic inhibition and regulation of Notch1 and HPSE CTC markers on BCBM onset; 2) link combinatorial Notch1/HPSE CTC subsets to clinical BCBM; 3) define roles of Merlin affecting BMSM CTC pathways and BCBM oncogenesis. This inter-institutional project by an inter-disciplinary and well-integrated team will study and validate new and specific CTC markers responsible for CTC-induced BCBM. This project is paradigm-shifting and has high therapeutic impact. We are uniquely positioned to perform this study not only for having access to an extensive cohort of blood samples and volume from patients diagnosed with or without BCBM but also for the availability of human CTC lines our laboratory was first to develop, and the extensive expertise combining multiple and complementary CTC platforms and approaches unlike other groups in the country.

 描述（由申请人提供）：乳腺癌脑转移（BCBM）是一种破坏性疾病，其机制尚未得到充分研究，在很大程度上仍然未知。例如，虽然人表皮生长因子受体1和2（EGFR和HER 2）是BCBM的高风险预测因子，但HER 2/EGFR靶向治疗已被证明对BCBM大多无效。尽管循环肿瘤细胞（CTC）作为难治性转移的“种子”的概念已经确立，但实际上对乳腺癌脑定殖CTC的性质和生物标志物一无所知。因此，迫切需要新的CTC靶生物标志物来对抗BCBM。我们已经发现了不表达上皮细胞粘附分子（EpCAM阴性CTC）并且在异种移植物中具有高BCBM能力的CTC。这些CTC不能被CellSearchTM检测到，CellSearchTM是一种FDA批准的预后测试，仅评估EpCAM阳性CTC。我们在EpCAM阴性CTC中发现了两种有希望的标记物，以预测临床中的BCBM：Notch 1，一种重要的肿瘤起始细胞生物标记物，和乙酰肝素酶（HPSE），一种有效的致瘤、血管生成和促转移分子。我们的结果表明，BCBM诱导的CTC具有“脑转移选择性标志物（BMSM）”特征或HER 2 +/EGFR+/Notch 1 +/HPSE+ CTC。因此，Notch 1和HPSE可以代表新的和特异性的CTC生物标志物。此外，我们在Notch 1/HPSE表达CTC中检测到肿瘤调节因子Merlin（2型神经纤维瘤病）的最高表达。通过PAK 1激酶磷酸化Merlin（P-S518 Merlin）导致Merlin的致癌功能，其与BMSM CTC途径及其调节相关。基于这些发现，我们的假设是Notch 1和HPSE是新的CTC生物标志物，可以预测原发性乳腺癌脑转移的存在;它们可以成为预防继发性BCBM的潜在治疗靶点。该提案的目的是进行临床前转化研究和靶点验证，以提名HPSE和Notch 1作为乳腺癌脑归巢CTC的关键生物标志物。我们将研究来自HER 2+患者的CTC，因为这种乳腺癌亚型发生BCBM的风险远高于平均水平。具体而言，我们将：1）确定Notch 1和HPSE CTC标志物的治疗性抑制和调节对BCBM发作的影响; 2）将组合Notch 1/HPSE CTC亚群与临床BCBM联系起来; 3）确定Merlin影响BMSM CTC途径和BCBM肿瘤发生的作用。由一个跨学科和综合性良好的团队开展的这一机构间项目将研究和验证负责CTC诱导的BCBM的新的和特定的CTC标志物。该项目是范式转变，具有很高的治疗效果。我们在进行这项研究方面具有独特的优势，不仅可以获得诊断为BCBM或无BCBM患者的大量血液样本和体积，而且还可以获得我们实验室首先开发的人类CTC细胞系，以及与该国其他团体不同的结合多种互补CTC平台和方法的广泛专业知识。

项目成果

Targeting USP7 Identifies a Metastasis-Competent State within Bone Marrow-Resident Melanoma CTCs.

• DOI: 10.1158/0008-5472.can-18-0644
• 发表时间: 2018-09-15
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: Vishnoi M;Boral D;Liu H;Sprouse ML;Yin W;Goswami-Sewell D;Tetzlaff MT;Davies MA;Oliva ICG;Marchetti D
• 通讯作者: Marchetti D

Molecular characterization of breast cancer CTCs associated with brain metastasis.

• DOI: 10.1038/s41467-017-00196-1
• 发表时间: 2017-08-04
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Boral D;Vishnoi M;Liu HN;Yin W;Sprouse ML;Scamardo A;Hong DS;Tan TZ;Thiery JP;Chang JC;Marchetti D
• 通讯作者: Marchetti D