Rapid and deterministic generation of microglia from human pluripotent stem cells - an in vitro platform for disease progression in neurodegeneration and neuroinflammation

从人类多能干细胞中快速、确定性地产生小胶质细胞——神经退行性变和神经炎症疾病进展的体外平台

基本信息

项目摘要

Microglia are the resident immune cells in the central nervous system (CNS) and implicated in the onset and progression of many neurological diseases. They originate from primitive yolk sac macrophages that colonise the developing CNS during early embryonic development. Once the embryonic microglial population is established, it is maintained throughout life by local proliferation, not replacement by bone-marrow-derived myeloid cells. Enormous research efforts are currently undertaken to establish faithful human microglia in vitro models as platform for disease modelling and drug discovery. Only most recently, the first protocols for the derivation of microglia-like cells from human pluripotent stem cells were reported. However, their long duration (35-80 days, depending on the protocol) and need for mechanical isolation steps to enrich certain developmental intermediate cell populations are likely to prohibit their widespread application. Here, I propose to establish a novel protocol for the generation of pure populations of microglia from human pluripotent stem cells at unprecedented speed and efficiency. I present preliminary data that this key step is feasible. Human microglia will be kept either in monoculture or placed in 2D coculture with human cortical neurons or 3D human brain organoids to establish a versatile human microglia platform. The different levels of complexity of the human microglia in vitro model will complement each other for diverse research questions ranging from reductionist studies in monoculture to studies of complex cellular interactions in an organoid setting. Microglia phenotypes are highly environment dependent. In the present application, we will apply the human microglia in vitro model to determine key cell-cell interactions that determine the human homeostatic microglia phenotype in brain organoids (as proxy of their in vivo counterparts) in contrast to the artificial in vitro phenotype. To this aim we will perform single cell RNA sequencing combined with the novel CITE-seq technology for simultaneous protein quantification. This will allow us to generate a comprehensive cellular interactome based on ligand receptor pair analysis. Finally, we will coculture microglia with tau-mutant cortical neurons and organoids to elicit and characterise a disease-associated microglia phenotype and unveil key cell-cell interactions that are associated with the phenotypic switch from homeostatic to disease-associated microglia.
小胶质细胞是中枢神经系统(CNS)中的常驻免疫细胞,并与许多神经系统疾病的发生和进展有关。它们起源于原始卵黄囊巨噬细胞,在早期胚胎发育期间定殖于发育中的CNS。一旦胚胎小胶质细胞群体建立,它在整个生命过程中通过局部增殖维持,而不是被骨髓来源的髓样细胞取代。目前正在进行大量的研究工作,以建立可靠的人类小胶质细胞体外模型,作为疾病建模和药物发现的平台。直到最近,才报道了从人类多能干细胞中衍生小胶质细胞样细胞的第一个方案。然而,它们的持续时间长(35-80天,取决于方案)并且需要机械分离步骤来富集某些发育中间细胞群,这可能会阻碍它们的广泛应用。在这里,我建议建立一个新的协议,以前所未有的速度和效率从人类多能干细胞中产生纯的小胶质细胞群体。我提供的初步数据表明这一关键步骤是可行的。人类小胶质细胞将保持在单一培养中或与人类皮质神经元或3D人脑类器官一起置于2D共培养中,以建立通用的人类小胶质细胞平台。人类小胶质细胞体外模型的不同复杂程度将相互补充,以解决各种研究问题,从单一培养中的还原论研究到类器官环境中复杂细胞相互作用的研究。小胶质细胞表型是高度环境依赖性的。在本申请中,我们将应用人小胶质细胞体外模型来确定关键的细胞-细胞相互作用,所述关键的细胞-细胞相互作用确定与人工体外表型相反的脑类器官中的人稳态小胶质细胞表型(作为其体内对应物的代表)。为此,我们将进行单细胞RNA测序,结合新的CITE-seq技术进行同时蛋白质定量。这将使我们能够基于配体受体对分析产生全面的细胞相互作用组。最后,我们将小胶质细胞与tau突变的皮质神经元和类器官共培养,以引发和抑制疾病相关的小胶质细胞表型,并揭示与从稳态到疾病相关的小胶质细胞的表型转换相关的关键细胞-细胞相互作用。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Privatdozent Dr. Matthias Pawlowski其他文献

Privatdozent Dr. Matthias Pawlowski的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('Privatdozent Dr. Matthias Pawlowski', 18)}}的其他基金

Induction of oligodendrocyte lineage cells by direct cellular reprogramming.
通过直接细胞重编程诱导少突胶质细胞系细胞。
  • 批准号:
    237496042
  • 财政年份:
    2013
  • 资助金额:
    --
  • 项目类别:
    Research Fellowships

相似海外基金

Deterministic quantum gate between photons in a next-generation light-matter interface
下一代光-物质界面中光子之间的确定性量子门
  • 批准号:
    EP/W035839/2
  • 财政年份:
    2024
  • 资助金额:
    --
  • 项目类别:
    Research Grant
Scalable Algorithms for Deterministic Global Optimization With Parallel Architectures
使用并行架构实现确定性全局优化的可扩展算法
  • 批准号:
    2330054
  • 财政年份:
    2024
  • 资助金额:
    --
  • 项目类别:
    Standard Grant
Collaborative Research: SaTC: CORE: Small: Understanding and Taming Deterministic Model Bit Flip attacks in Deep Neural Networks
协作研究:SaTC:核心:小型:理解和驯服深度神经网络中的确定性模型位翻转攻击
  • 批准号:
    2342618
  • 财政年份:
    2023
  • 资助金额:
    --
  • 项目类别:
    Standard Grant
Mean-Field and Singular Limits of Deterministic and Stochastic Interacting Particle Systems
确定性和随机相互作用粒子系统的平均场和奇异极限
  • 批准号:
    2345533
  • 财政年份:
    2023
  • 资助金额:
    --
  • 项目类别:
    Standard Grant
Methods for deterministic treatment effect estimates for clinical trials with missing data
缺失数据的临床试验的确定性治疗效果估计方法
  • 批准号:
    2886293
  • 财政年份:
    2023
  • 资助金额:
    --
  • 项目类别:
    Studentship
Towards deterministic atomic scale manufacturing of next-generation quantum devices
迈向下一代量子器件的确定性原子尺度制造
  • 批准号:
    EP/X021963/1
  • 财政年份:
    2023
  • 资助金额:
    --
  • 项目类别:
    Fellowship
Application of deterministic dopant devices to probabilistic information processing, quantum computing/measurement
确定性掺杂器件在概率信息处理、量子计算/测量中的应用
  • 批准号:
    23H00169
  • 财政年份:
    2023
  • 资助金额:
    --
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Deterministic quantum gate between photons in a next-generation light-matter interface
下一代光-物质界面中光子之间的确定性量子门
  • 批准号:
    EP/W035839/1
  • 财政年份:
    2023
  • 资助金额:
    --
  • 项目类别:
    Research Grant
CloudEnergyBalance: Simple climate models to quantify impact of large-scale cloudiness & deterministic chaos on climatic variability & tipping points
CloudEnergyBalance:用于量化大规模多云影响的简单气候模型
  • 批准号:
    EP/Y01653X/1
  • 财政年份:
    2023
  • 资助金额:
    --
  • 项目类别:
    Fellowship
PRogrammable AI-Enabled DeterminIstiC neTworking for 6G
适用于 6G 的可编程 AI 确定性网络
  • 批准号:
    10060071
  • 财政年份:
    2023
  • 资助金额:
    --
  • 项目类别:
    EU-Funded
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了