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

用于临床研究中单细胞基因组学的可部署一次性设备

基本信息

批准号：
10258377
负责人：
Todd M Gierahn
金额：
$ 102.55万
依托单位：
HONEYCOMB BIOTECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10258377
关键词：
Agreement Bar Codes Benchmarking Biological Sciences Biotechnology Blood Blood specimen Bone Marrow COVID-19 pandemic Cell Membrane Permeability Cells Cellular Assay Cerebrospinal Fluid Clinical Clinical Research Clinical Trials Collection Complex Cryopreservation Data Data Set Development Devices Disease Employee Equipment Functional disorder Gene Expression Gene Expression Profile Genomics Goals Health Human Knowledge Lead Libraries Life Logistics Love Manuals Membrane Modernization Monitor Performance Pharmacologic Substance Pharmacotherapy Phase Population Process Production Protocols documentation Quality Control RNA Reagent Recovery Research Sampling Senior Scientist Symptoms System Technology Testing Time Training Transcript Vaccine Clinical Trial Validation Veterans Virus Work alpha-SNAP base clinical care clinical research site data quality design elastomeric experience instrumentation macromolecule manufacturing process portability predict clinical outcome preservation product development sample collection seal single cell analysis single cell technology single-cell RNA sequencing tool uptake

项目摘要

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.

项目概要单细胞 RNA-seq (scRNA-seq) 已成为描述细胞状态的有效工具异质细胞群。然而，在临床研究和由于样本后不久需要稳定转录图谱的后勤工作，护理工作具有挑战性收藏。我们已经证明了集成样本收集的一次性盒的可行性，将临床样本存储和处理至 scRNA-seq 文库。该盒利用皮升井阵列预加载有条形码的转录捕获珠，用于收集和分离单细胞。一个细胞将保存试剂添加到盒中以将转录物谱稳定在其天然状态。一个弹性体柱塞用于通过简单的卡扣将阵列与半透膜密封机制。密封的盒可以发送到中央设施进行批量处理到测序图书馆。在临床现场收集和储存细胞不需要专门的设备，从而能够将该技术轻松部署到多个临床站点。我们建议完成开发并验证盒式磁带的制造和用户工作流程。具体来说，我们将 1) 强化用于生成包埋盒并执行多批次验证活动的制造协议，2) 开发并验证一个简单的工作流程，用于处理临床样本并将其加载到盒中， 3) 验证分布式实验室对收集和存储在盒中的样本的中央处理。到为了实现这些目标，我们创建了一个强大的联盟，包括核心 Honeycomb 团队、顾问 Alex Shalek 博士和 Christopher Love 博士是两位单细胞分析专家，也是该技术的共同开发者基础技术，以及两家在该领域拥有丰富经验的大型制药合作伙伴临床研究的要求，并将广泛验证该技术。