High quality CTC isolation using microbubbles for downstream molecular analysis
使用微泡进行高质量 CTC 分离以进行下游分子分析
基本信息
- 批准号:8737211
- 负责人:
- 金额:$ 30.88万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2013
- 资助国家:美国
- 起止时间:2013-09-18 至 2016-08-31
- 项目状态:已结题
- 来源:
- 关键词:AntibodiesAwardBenchmarkingBiologicalBiological MarkersBiopsyBloodBlood specimenBody FluidsBrain NeoplasmsCancer InterventionCancer PrognosisCell SeparationCellsCentrifugationClinical ManagementClipCollecting CellComplexDiagnosisDiagnosticFluorocarbonsGoalsHarvestHuman ResourcesIn VitroLeukocytesLipidsLiquid substanceMalignant NeoplasmsMalignant neoplasm of brainMethodsMicrobubblesMolecularMolecular AnalysisNeoplasm Circulating CellsPatientsPhasePrimary NeoplasmProceduresProcessRNA DegradationRecurrent tumorSamplingScreening for cancerSensitivity and SpecificitySourceSpecificitySpecimenSpeedSystemTACSTD2 geneTechniquesTechnologyTestingTimeTissuesbasecancer cellcell growthcost effectivedrug sensitivityflexibilitygenetic profilingimprovedinnovationneoplastic cellnew technologyoutcome forecastprotein degradationpublic health relevancetreatment planningtumor progression
项目摘要
DESCRIPTION (provided by applicant): Early cancer detection and intervention are crucial for long-term survival of either primary or recurrent tumors. Circulating tumor cells (CTCs) could be isolated for the diagnosis, prognosis, and treatment planning of cancer. Genetic profiling and expression analysis of CTC could provide additional invaluable information for accurate diagnosis and prognosis. Moreover, in vitro culture of CTC can be applied for functional analysis of cancer cells, such as drug sensitivity test. To achieve this goal, a transformative technology must be able to unmistakably isolate a few cancer cells using cost-effective and non-invasive procedures. Under the previous R21 IMAT award, we developed a novel technology for rare tumor cell isolation. Lipid shell-perfluorocarbon buoyant microbubbles (MBs) coated with anti-EpCAM antibody immediately attached to tumor cells in unfractionated blood, and isolated the cells after a quick centrifugation step (Shi et al., PLoS One, 2013). A short video clip to demonstrate this technology is included in this application. In conclusion, MB technique offers unique advantages over the existing immuno-enrichment technologies: (a) Short processing time can avoid RNA and protein degradation; (b) Scalability (isolation of cells from large volume sample) can help obtain large numbers of CTCs; (c) Specificity (minimal carryover of leukocytes) can improve sensitivity and specificity of molecular analysis; (d) Flexibility (cells ae collected in a very concentrated volume of 5-10 ¿l and could be used for cell growth, immunostaining and/or molecular analysis. The R33 phase of this proposal is focused to further develop MB isolation method to achieve high purity isolation of CTCs from metastatic brain cancers for enumeration and molecular and functional analyses. We set the following specific aims of the study: (1) Optimize MB method to achieve high speed, high purity harvesting of tumor cells spiked in 7-20ml blood sample; (2) Applications of MB isolated CTCs for PCR analysis and culturing; (3) Validate the MB method for sensitivity and specificity of CTC enumeration from blood of metastatic brain tumor patients using CellSearch system as a benchmark. Simple but robust CTC isolation technologies that result in high quality CTC sample for downstream analysis will have transformative impact on clinical management of malignancies and early diagnostics of asymptomatic cancers.
描述(由申请人提供):早期癌症检测和干预对于原发性或复发性肿瘤的长期生存至关重要。循环肿瘤细胞(CTC)可以被分离用于癌症的诊断、预后和治疗计划。CTC的基因表达谱分析可为准确诊断和预后提供额外的宝贵信息。此外,CTC的体外培养可用于癌细胞的功能分析,例如药物敏感性测试。为了实现这一目标,变革性技术必须能够使用具有成本效益的非侵入性程序明确地分离出一些癌细胞。 在之前的R21 IMAT奖下,我们开发了一种用于罕见肿瘤细胞分离的新技术。用抗EpCAM抗体包被的脂质壳-全氟化碳浮力微泡(MB)立即附着于未分级血液中的肿瘤细胞,并在快速离心步骤后分离细胞(Shi等人,PLoS One,2013)。一个简短的视频剪辑,以证明这项技术是包括在这个应用程序。总之,MB技术提供了优于现有免疫富集技术的独特优势:(a)短的处理时间可以避免RNA和蛋白质降解;(B)可扩展性(从大体积样品中分离细胞)可以帮助获得大量的CTC;(白细胞的最小携带)可以提高分子分析的灵敏度和特异性;(d)灵活性(细胞收集在5-10升的非常浓缩的体积中,可用于细胞生长、免疫染色和/或分子分析)。 该提案的R33阶段重点是进一步开发MB分离方法,以实现从转移性脑癌中高纯度分离CTC,用于计数以及分子和功能分析。我们设定了以下具体研究目标:(1)优化MB方法,以实现7- 20 ml血液样品中掺入的肿瘤细胞的高速、高纯度收获;(2)MB分离的CTC用于PCR分析和培养;(3)以CellSearch系统为基准,验证MB方法对转移性脑肿瘤患者血液中CTC计数的灵敏度和特异性。 简单但强大的CTC分离技术,导致高质量的CTC样品用于下游分析,将对恶性肿瘤的临床管理和无症状癌症的早期诊断产生变革性影响。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Yu-Tsueng Liu其他文献
Yu-Tsueng Liu的其他文献
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{{ truncateString('Yu-Tsueng Liu', 18)}}的其他基金
High quality CTC isolation using microbubbles for downstream molecular analysis
使用微泡进行高质量 CTC 分离以进行下游分子分析
- 批准号:
8915098 - 财政年份:2013
- 资助金额:
$ 30.88万 - 项目类别:
High quality CTC isolation using microbubbles for downstream molecular analysis
使用微泡进行高质量 CTC 分离以进行下游分子分析
- 批准号:
8624981 - 财政年份:2013
- 资助金额:
$ 30.88万 - 项目类别:
Non-invasive sampling of DNA markers for pancreatic cancer screening
用于胰腺癌筛查的 DNA 标记物无创采样
- 批准号:
8435334 - 财政年份:2012
- 资助金额:
$ 30.88万 - 项目类别:
Prostate Fusion Gene Variants as a Cancer Biomarker
前列腺融合基因变异体作为癌症生物标志物
- 批准号:
7586685 - 财政年份:2008
- 资助金额:
$ 30.88万 - 项目类别:
Prostate Fusion Gene Variants as a Cancer Biomarker
前列腺融合基因变异体作为癌症生物标志物
- 批准号:
7450700 - 财政年份:2008
- 资助金额:
$ 30.88万 - 项目类别:
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