Cell free RNA liquid biopsies

摘要 本提案的最终目标是验证RNA液体活检平台并将其过渡到具有特定实用性 用于造血骨髓（BM）小生境的研究和非侵入性评估。除了它们 依赖于从垂死的患病细胞分泌的物质，无细胞DNA（cfDNA）平台主要限于 检测体细胞基因组序列、拷贝数或甲基化状态的变化。许多生物和 临床事件，如造血、纤维化和肿瘤发生，通过全局变化执行， 转录调控我的博士生导师丹尼尔金博士和我建立了一个平台， 在人类血浆中的活检已经证明了在这两个方面的显着诊断和监测潜力， 胰腺癌和新冠肺炎患者。我建议继续这项研究，以完成我的博士学位， 评估exRNA平台在60例肺腺癌新队列中的诊断性能 （LUAD）患者与匹配对照。此外，我将运用我在exRNA液体活检方面的专业知识， 测定骨髓中造血和造血干细胞（HSC）的转录动力学 (BM)利基目前，骨髓活组织检查或抽吸用于确定骨髓健康， 生产这些技术是昂贵的，并且需要对受试者进行镇静和/或疼痛缓解， 可能导致长期不适、感染、出血过多和其他副作用。BM愿望 仍然是HSC移植恢复、白血病和淋巴瘤、血液病和淋巴瘤的重要诊断和监测工具。 细胞病理学和未知来源的感染，但主要读出仍然是识别和计数， 细胞类型。我假设骨髓内的造血谱系分泌反映细胞状态的exRNA， 可用于非侵入性RNA液体活检以详细研究转录和群体的身份 在BM。我建议鉴定HSC表达和分泌的exRNA，以及其余的exRNA。 以开发一个平台，将外周血去卷积成造血谱系的组分 造血细胞类型，而不需要HSC动员。

ABSTRACT The ultimate goal of this proposal is to verify an RNA liquid biopsy platform and transition it to have specific utility for the study, and non-invasive assessment, of the hematopoietic bone marrow (BM) niche. In addition to their reliance of material excreted from dying, diseased cells, cell-free DNA (cfDNA) platforms are limited to primarily detecting changes in somatic genomic sequence, copy number, or methylation status. Many biological and clinical events, such as hematopoiesis, fibrosis, and tumorigenesis, are executed via global changes in transcriptional regulation. My PhD advisor Dr. Daniel Kim and I have established a platform for exRNA liquid biopsy in human blood-plasma that has demonstrated significant diagnostic and monitoring potential in both Pancreatic cancer and COVID-19 patients. I propose to continue this research to complete my PhD by assessing diagnostic performance of the exRNA platform in a new cohort of 60 lung adenocarcinoma (LUAD) patients with matched controls. Furthermore, I will apply my expertise in exRNA liquid biopsies to assay the transcriptional dynamics of hematopoiesis and hematopoietic stem cells (HSCs) in the bone marrow (BM) niche. Currently, bone marrow biopsies or aspirations are used to determine bone marrow health and production. These techniques are costly and require sedation and/or pain relief for the subject and have the potential to lead to long term discomfort, infection, excessive bleeding, and other side-effects. BM aspirations remain a critical diagnostic and monitoring tool for HSC transplant recovery, leukemias and lymphomas, blood cell pathologies, and infections of unknown origin but the primary readout remains identification and counting of cell types. I hypothesize that hematopoietic lineages within the BM secrete exRNA that reflect cell state and identity that can be used in a non-invasive RNA liquid biopsy for detailed study of transcription and populations within the BM. I propose to identify exRNA expressed and secreted by HSCs and the remaining hematopoietic lineage in order to develop a platform to deconvolute peripheral blood into constituent hematopoietic cell types without the need for HSC mobilization.