Scanning for intensely fluorescent targets (SIFT): a new tool to study pathological protein aggregate formation in MFM and its reversal through pharmacologic intervention

我们的目的是阐明由于MFM基因突变导致的肌原纤维肌病中导致病理性蛋白质聚集和肌肉病理的机制。作为一种新的方法，我们将使用多色共聚焦单粒子光谱分析方法，如扫描强烈荧光靶标(SIFT)来分析结蛋白突变体和其他MFM相关蛋白的蛋白质聚集体，这些蛋白质聚集体以前是通过转染和体外组装研究来表征的。重点将集中在病理性蛋白质聚集体的形成、分子结构以及活细胞和单分子水平上的同源和异源蛋白质-蛋白质相互作用方面的特征。治疗的前景是通过测试直接针对病理性蛋白寡聚体、等位基因特异性击倒、热休克蛋白诱导剂以及通过自噬和蛋白酶体代谢影响聚集清除的化合物来解决的。

We aim to elucidate the mechanisms leading to pathological protein aggregation and muscle pathology in myofibrillar myopathies due to mutations in MFM causing genes. As a novel approach, we will use multi-color confocal single-particle spectroscopy methods such as scanning for intensely fluorescing targets (SIFT) to analyze protein aggregates of desmin mutants and other MFM-associated proteins, which have previously been characterized by transfection and in vitro assembly studies. The focus will be the characterization of pathological protein aggregates in regard to formation, molecular architecture, and homologeous as well as heterologeous protein-protein interactions in living cells and at the single-molecule level. Therapeutic perspectives are addressed by testing novel compounds directly targeting pathological protein oligomers, allelle-specific knock-down, heat shock protein inducers, and compounds affecting aggregate clearance by autophagy and proteasomal metabolism.