Clinical development of a blood based assay that detects the BRAF V600E mutation in melanoma and thyroid cancer patients

项目总结/摘要 大约一半的恶性黑色素瘤和乳头状甲状腺癌（PTC）患者有一个恶性肿瘤。 携带BRAFV 600突变的肿瘤。突变检测是确定基因突变的关键分支点 BRAF靶向治疗是否是晚期（不可切除的III期和IV期）患者的一种选择 黑素瘤此外，知道PTC患者的BRAF状态可以是临床上可行的，因为它可以指导患者的BRAF状态。 初次手术的范围（肺叶切除术与甲状腺全切除术，以及考虑中央 淋巴结切除术），监测成像方法（放射性碘扫描与PET-CT），以及 辅助治疗目前，商业化的BRAF突变分析测定受到灵敏度的限制， 在大量接受BRAF抑制剂治疗或以下治疗的患者中缺乏可靠的临床验证 在高危环境下进行肿瘤切除随着BRAF靶向治疗现已成为标准 对于BRAF突变型恶性肿瘤患者的治疗，我们认为高敏感性， 基于血液的测定具有极大地改善患者护理的潜力。我们以前 描述了用高灵敏度测定法进行的测试，该测定法能够检测在人外周血中的BRAFV 600突变。 血液与BRAF突变黑色素瘤患者，现在已经表明，这种测定具有良好的， 在黑色素瘤和PTC患者中相对维斯基于组织的分析的灵敏度和特异性。在这 我们的目标是优化我们在贝斯以色列女执事医疗研究实验室的测定 在UH 2部分期间， 在马萨诸塞州总医院的CLIA批准的实验室批准并协调该测定 （MGH）.一旦检测试剂盒经过优化、协调并成功转移至CLIA， 在MGH批准的实验室，我们计划在III期和III期患者中临床验证我们的检测方法 IV黑色素瘤和可切除和转移性PTC，通过分析从 几个不同的临床试验这些包括达拉非尼加曲美替尼的新辅助试验， III期BRAF突变型黑色素瘤患者（NCT 02231775），一项随机III期研究， 达拉非尼+曲美替尼，然后是伊匹单抗+纳武单抗的顺序，与其他 在初治晚期黑色素瘤患者中的序列（NCT 02224781），III期 随机双盲研究，比较4周疗程后的完全缓解率， MEK抑制剂、司美替尼或安慰剂与单剂量放射性碘辅助治疗 两种II期靶向治疗，一种是 多酪氨酸激酶抑制剂乐伐替尼（NCT 02657369），另一种与mTOR抑制剂 MLN0128（NCT 02244463）。 分化型甲状腺癌（NCT 02393690），和

Project Summary/Abstract Approximately half of patients with malignant melanoma and papillary thyroid cancer (PTC) have a tumor that harbors a BRAFV600 mutation. Mutation detection is the critical branch-point in determining whether BRAF-targeted therapy is an option for patients with advanced (unresectable Stage III and IV) melanoma. Also, knowing a PTC patient's BRAF status may be clinically actionable, since it may guide the extent of initial surgery (lobectomy versus total thyroidectomy and consideration of central lymphadenectomy), the approach to surveillance imaging (radioactive iodine scan versus PET-CT), and adjuvant therapy. Currently, commercial BRAF mutational analysis assays are limited by sensitivity and lack robust clinical validation in large numbers of patients treated with a BRAF inhibitor or following tumor resection in the high-risk setting. With BRAF targeted therapy now established as a standard therapy for patients with BRAF mutant malignancies, we feel that the development of highly-sensitive, blood-based assays have the potential to greatly improve the care of patients. We have formerly described testing with a highly sensitive assay that has the ability to detect BRAFV600 mutations in the blood of patients with BRAF mutant melanoma, and now have shown that this assay has excellent sensitivity and specificity vis-à-vis tissue based analysis in both melanoma and PTC patients. In this proposal, we aim to optimize our assay in the research laboratory at Beth Israel Deaconess Medical Center, where all the previous work with the assay has been performed, during the UH2 portion of the grant and harmonize the assay in a CLIA-approved laboratory at Massachusetts General Hospital (MGH). Once the assay has been optimized, harmonized and successfully transferred to the CLIA- approved laboratory at MGH, we plan to clinically validate our assay in patients with Stage III and Stage IV melanoma and resectable and metastatic PTC through the analysis of samples obtained from several different clinical trials. These include a neoadjuvant trial of dabrafenib plus trametinib in patients with Stage III BRAF-mutant melanoma (NCT02231775), a randomized Phase III study of the sequence of dabrafenib plus trametinib followed by ipilimumab plus nivolumab compared with the other sequence in treatment naïve patients with advanced melanoma (NCT02224781), a Phase III randomized double blind study comparing the complete remission rate following a 4-week course of the MEK inhibitor, selumetinib, or placebo and a single dose adjuvant radioactive iodine therapy in patients two phase II targeted therapies, one with the multi-tyrosine kinase inhibitor lenvatinib (NCT02657369) and the other with the mTOR inhibitor MLN0128 (NCT02244463). with differentiated thyroid cancer (NCT02393690), and