3D co-culture system to characterize and target leukemia stem cells

项目概要： 造血干细胞（HSC）存在于骨髓（BM）中，并且它们的功能由它们的细胞周期调控。 与BM微环境的相互作用。白血病干细胞（LSC）已被证明能够 引发和维持急性髓性白血病（AML）和急性淋巴细胞白血病（ALL）。此外，委员会认为， LSC与其微环境的相互作用已被证明在抗性中起关键作用 LSC的治疗。研究LSCs的传统方法和抗白血病药物对其的影响 2D单层细胞培养。随着复杂的细胞-基质相互作用的不断阐明， 很明显，这个模型不再是BM利基复杂性的充分代表。因此 开发一个更好地模拟这种微环境的模型是必要的，既要研究LSC特性， 以及用于在反映微环境贡献的系统中测试药物效果。我们 假设能够重现BM生态位条件以维持LSC的培养系统将允许 为了更好地了解它们的生物学和发现新的治疗方法， BM中的LSC。我们建议实施新的方法来评估BM-LSC的相互作用，以改善 以患者特异性方式评估LSC的功能、耐药性和干细胞特性。我们 初步数据表明3D球状体（类器官结构）可以概括BM生态位特征， 例如缺氧。此外，我们还观察到，LSC得到了更好的维护， 与更常规的培养系统相比，使用3D基质方法的干性特性。因此，在本发明中， 我们建议评估AML（具有良好特征的突变谱），以确定其存活率 生态位相互作用背景下的依赖关系。该项目的最终目标是确定新的治疗方法， 直接或通过破坏BM-LSC保护性相互作用靶向LSC的方法。

PROJECT SUMMARY: Hematopoietic stem cells (HSCs) reside in the bone marrow (BM) and their function is regulated by their interactions with the BM microenvironment. Leukemia stem cells (LSCs) have been shown to be capable of initiating and maintaining acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). Furthermore, the interactions of LSCs with their microenvironment have been shown to play a critical role in the resistance of LSCs to therapy. The traditional approach of investigating LSCs and anti- leukemic agents’ effects on them has been the 2D monolayer cell culture. As complex cell-matrix interactions continue to be elucidated, it is clear that this model is no longer a sufficient representative of the intricacies of the BM niche. Thus, the development of a model that better mimics this microenvironment is imperative, both to study LSC properties, as well as for testing drug effects in a system that reflects the contributions of the microenvironment. We hypothesize that a culture system capable of recapitulating the BM niche conditions to maintain LSCs will allow for the better understanding of their biology and the discovery of novel therapeutic approaches that can act on LSCs in the BM. We propose to implement novel approaches to evaluate BM-LSC interactions to improve the assessment of the function, drug resistance and stem cell properties of LSCs in a patient-specific manner. Our preliminary data suggest that the 3D spheroids (organoid-like structures) can recapitulate BM niche features, such as hypoxia. In addition, we have observed that the LSCs are better maintained and retain functional stemness properties using the 3D stromal approach compared to more conventional culture systems. Thus, we propose to evaluate AML (with a well-characterized mutational profile) to determine their survival dependencies in the context of niche interactions. The ultimate goal of this project is to identify new therapeutic approaches to target LSCs either directly or through disruption of the BM-LSC protective interactions.