Detecting Cancer at the Single-Cell Level Using Endogenous Signal Biomolecular Sensors

癌症是一类以不受控制的生长、侵袭和转移为特征的细胞疾病。虽然癌症是多方面的，具有复杂的潜在遗传决定因素，但新出现的证据表明，在不同组织和癌症类型的细胞内信号传导水平上存在共同的特征。能够检测细胞中这些细胞内分子信号的组合的系统对于在单细胞水平上诊断、预防、治疗和监测疾病具有显著的潜力。最近，科学家们已经开发出第一代遗传电路和传感器，它们在细胞中作为信息处理系统运行。拟议的研究将开发和研究分子生物传感器，旨在检测和监测内源性微小RNA，这是一类通过靶向RNA转录物来调节基因表达的小型非编码RNA。本研究的目标是：（a）设计和优化对内源性microRNA组合具有灵敏度和特异性的分子生物传感器;（B）开发基于NCI-60面板的具有集成生物传感器的单克隆肿瘤细胞系;（c）通过监测microRNA在细胞周期中的表达来研究其与疾病的关系。MicroRNA已成为发育和疾病（包括癌症）的重要调节剂。实验证据表明，在大多数人类恶性肿瘤中存在异常的microRNA表达谱。可靠地处理microRNA信息（单细胞分辨率）的能力将为基础癌症研究提供新的工具，并导致新的癌症治疗模式。具体来说，microRNA生物传感器在肿瘤细胞系中的应用将极大地促进我们对活细胞中microRNA表达和动力学的理解。使用选定的microRNA传感器开发单克隆细胞系将为一类新的microRNA研究工具铺平道路。这将是一个重要的新方向，将引发其他细胞系和其他microRNA的一系列等效研究。更广泛的影响：实验将在NCI-60结肠、血液、肺和胰腺肿瘤细胞系中进行。选择特定细胞系的标准基于：美国肺癌和结肠癌的患病率（分别是男性和女性癌症死亡的主要原因，以及男性和女性癌症死亡的第二大原因）;因为胰腺癌和血液癌的5年生存率很低，（特别是胰腺癌）的发展强调了对新的诊断和治疗工具的迫切需要。将向科学界提供含有生物分子网络的单克隆细胞系的集合。拟议的研究还将形成一个教育平台，学生将接触到一个综合的研究环境，在生物学和工程学的交叉点结合理论和实验。

Cancer is a class of cellular diseases characterized by uncontrolled growth, invasion, and metastasis. Although cancer is multifaceted with complex underlying genetic determinants, emerging evidence points to common signatures at the level of intracellular signaling across different tissue and cancer types. A system capable of detecting combinations of these intracellular molecular signals in cells has significant potential for the diagnosis, prevention, treatment, and monitoring of the disease at a single-cell level. Recently, scientists have developed first generation genetic circuits and sensors that operate as information processing systems in cells. The proposed research will develop and study molecular biosensors engineered to detect and monitor endogenous microRNAs, a class of small, noncoding RNAs that regulate gene expression by targeting RNA transcripts. The objectives of the proposed research are to: (a) design and optimize molecular biosensors that respond with sensitivity and specificity to combinations of endogenous microRNAs, (b) develop monoclonal tumor cell lines with integrated biosensors based on the NCI-60 panel, and (c) study microRNAs and their relationship to disease by monitoring their expression through the cell cycle in response to drug treatment.Intellectual merit: MicroRNAs have emerged as important modulators in both development and disease, including cancer. Experimental evidence implicates aberrant microRNA expression profiles in most human malignancies. The ability to process microRNA information reliably (with single-cell resolution) will generate new tools for basic cancer research and lead to new cancer treatment modalities. Specifically, the application of the microRNA biosensors to tumor cell lines will dramatically advance our understanding of microRNA expression and dynamics in living cells. The development of monoclonal cell lines with the selected microRNA sensors will pave the way for a new class of tools for the study of microRNAs. This will be a major new direction that will spark a range of equivalent studies in other cell lines and with other microRNAs. Broader impact: The experiments will be performed in NCI-60 colon, hematologic, lung, and pancreatic tumor cell lines. The criteria for the selection of the particular cell lines was based: on the prevalence of lung cancer and colon cancer in the United States (the leading cause of cancer death for both men and women and the second leading cause of cancer deaths for both men and women combined, respectively); because pancreatic and hematologic cancers have very poor 5-year survival (pancreatic cancer in particular) underscoring the urgent need for new diagnostic and treatment tools. The collection of monoclonal cell lines harboring the biomolecular networks will be made available to the scientific community. The proposed research will also form an educational platform where students will be exposed to an integrated research environment, combining theory and experiments at the intersection of biology and engineering.