Deployable, Single-use Device for Single Cell Genomics in Clinical Research

项目摘要 单细胞rna-seq(scrna-seq)已成为描述细胞状态的有效工具。 异质细胞群体。然而，利用这一强大的技术在临床研究和 CARE是具有挑战性的，因为在样本后不久稳定转录图谱的后勤工作 收集。我们已经证明了集成样本采集的一次性使用盒式磁带的可行性， 临床样本到scRNA-seq文库的存储和处理。这个盒式磁带有一个皮升的杠杆 阵列预加载条形码记录捕获珠，以收集和分离单个细胞。一间牢房 将保存剂添加到盒式磁带中，以使转录谱稳定在其自然状态。一个 使用弹性柱塞使用简单的卡扣将阵列与半透膜密封 机制。密封的盒式磁带可以被送到中央设施进行批处理以进行排序 图书馆。不需要专门的设备在临床现场收集和存储细胞，从而实现 将该技术轻松部署到多个临床站点。我们建议完成开发和 验证盒式磁带的制造和用户工作流。具体地说，我们将1)强化 用于产生盒式磁带和执行多批次验证活动的制造协议，2) 开发和验证用于处理临床样本并将其装入试剂盒的简单工作流程 3)验证分布式实验室收集和存储在盒式磁带中的样本的中央处理。至 为了实现这些目标，我们创建了一个强大的财团，包括核心蜂巢团队、顾问 Alex Shalek博士和Christopher Love博士，两位单细胞分析专家和 基础技术和两个大型制药合作伙伴，他们在 临床研究的要求，并将广泛验证该技术。

Project Summary Single cell RNA-seq (scRNA-seq) has emerged as a potent tool for delineating cell states in heterogeneous cell populations. However, leveraging this potent technology in clinical research and care is challenging due to the logistics of stabilizing the transcriptional profiles soon after sample collection. We have demonstrated feasibility of a single use cassette that integrates sample collection, storage and processing of clinical samples to scRNA-seq libraries. The cassette leverages a picoliter-well array pre-loaded with barcoded transcript capture beads to collect and isolate single cells. A cell preservation reagent is added to the cassette to stabilize the transcript profiles in their native state. An elastomeric plunger is used to seal the array with a semipermeable membrane using a simple snapping mechanism. The sealed cassette can be sent to a central facility for batch processing to sequencing libraries. No specialized equipment is required for collection and storing cells at clinical sites, enabling facile deployment of the technology to multiple clinical sites. We propose to finish development and validate the manufacturing and user workflows for the cassette. Specifically we will 1) harden the manufacturing protocols for generating the cassettes and perform a multi-lot validation campaign, 2) develop and validate a simple workflow for processing and loading clinical samples into the cassette and 3) validate central processing of samples collected and stored in the cassettes by distributed labs. To achieve these goals, we have created a strong consortium including the core Honeycomb team, advisors Dr. Alex Shalek and Dr. Christopher Love, two experts in single cell analysis and co-developers of the underlying technology, and two large pharma partners who have significant experience in the requirements for clinical research and will be extensively validating the technology.