Determinants of Cell State Reprogramming

细胞状态重编程的决定因素 项目概要 转录因子 (TF) 的强制表达可以重新编程细胞状态。直接重编程的发现 一直是我们理解细胞状态的分子和遗传驱动因素的催化剂。然而，尽管其 成功，仅确定了少数成功的重编程鸡尾酒，转换过程 通常效率低下，并且重编程失败的机制原因通常不清楚。因此，一个关键障碍 取得进一步进展的是我们对细胞状态转换的决定因素和机制的不完全理解。 我们的长期目标是了解细胞状态的遗传和分子基础，这具有重要意义 用于再生医学细胞状态的综合控制。为了实现这一目标，本提案的目标是 阐明细胞状态重编程的分子和细胞决定因素。具体来说，我们寻求解决 细胞状态重编程知识与三个中心研究方向的几个基本差距：1） 细胞重编程的表观遗传决定因素，2) 细胞的遗传和时间决定因素 分化，以及 3) 重编程的初始细胞状态决定因素。这个范围是通过我们的 将多因素、汇集扰动与每个细胞的多维读数相结合的创新平台 涵盖扰动（诱导转录因子）、机制（表观基因组）和表型（转录组）。通过追踪 数以千计的单个细胞正在跨越多种不同的扰动进行重新编程，我们的高 吞吐量功能实验将揭示见解并提出决定因素的机制 重新编程。我们的理由是，学习这些重新编程的基本规则将提高我们的能力 了解细胞状态的分子基础并改进操纵它的方法。

Determinants of cell state reprogramming PROJECT SUMMARY Forced expression of transcription factors (TFs) can reprogram cell state. The discovery of direct reprogramming has been a catalyst for our understanding of the molecular and genetic drivers of cell state. However, despite its successes, only a handful of successful reprogramming cocktails have been identified, the conversion process is often inefficient, and the mechanistic reasons for reprogramming failure are often unclear. Thus, a key obstacle to further progress is our incomplete understanding of the determinants and mechanisms of cell state conversion. Our long term goal is to understand the genetic and molecular basis of cell state, which has important implications for synthetic control of cell state for regenerative medicine. Towards this goal, the objective of this proposal is to elucidate the molecular and cellular determinants of cell state reprogramming. Specifically, we seek to address several fundamental gaps in knowledge on cell state reprogramming with three central research directions: 1) Epigenetic determinants of cellular reprogramming, 2) Genetic and temporal determinants of cellular differentiation, and 3) Initial cell state determinants on reprogramming. This scope is made possible by our innovative platform that couples multifactorial, pooled perturbation with multidimensional readouts for each cell spanning perturbations (induced TFs), mechanisms (epigenome), and phenotypes (transcriptome). By tracing thousands of individual cells on their path to reprogramming across a multitude of distinct perturbations, our high throughput functional experiments will reveal insights and suggest mechanisms for the determinants of reprogramming. Our rationale is that learning these fundamental rules of reprogramming will improve our understanding of the molecular basis of cell state and enable improved approaches to manipulate it.