Cell free RNA liquid biopsies

摘要 这项提议的最终目标是验证RNA液体活组织检查平台并将其转换为具有特定用途的平台 用于造血骨髓(BM)生态位的研究和非侵入性评估。除了他们的 依赖于死亡、患病细胞排出的物质，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、纤维化和肿瘤形成，是通过 转录调控。我的博士导师Daniel Kim博士和我已经建立了一个exRNA液体平台 在人类血浆中的活组织检查已经显示出对这两种疾病具有显著的诊断和监测潜力 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例新的肺腺癌队列中的诊断性能 (LUAD)患者与匹配的对照组。此外，我将运用我在exRNA液体活组织检查方面的专业知识 骨髓中造血和造血干细胞转录动态的检测 (BM)利基。目前，骨髓活检或活检被用来确定骨髓健康和 制作。这些技术成本高昂，需要对受试者进行镇静和/或止痛，并具有 可能导致长期不适、感染、出血过多和其他副作用。BM抱负 仍然是造血干细胞移植恢复、白血病和淋巴瘤、血液等的关键诊断和监测工具 细胞病理和来源不明的感染，但主要读数仍为鉴定和计数 单元类型。我假设骨髓中的造血系分泌exRNA来反映细胞状态和 可用于非侵入性RNA液体活检以详细研究转录和种群的身份 在BM内部。我建议鉴定由肝星状细胞表达和分泌的exRNA以及其余的 造血谱系为了开发一个平台将外周血去卷曲成成分 不需要动员HSC的造血细胞类型。

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.