There is growing evidence that macroautophagic cargo is not limited to bulk cytosol in response to starvation, and can occur selectively for substrates including aggregated proteins. It remains unclear, however, if starvation-induced and selective macroautophagy share identical adapter molecules to capture their cargo. Here we report that Alfy, a phosphatidylinositol 3-phosphate binding protein, is central to the selective elimination of aggregated proteins. We report that the loss of Alfy inhibits the clearance of inclusions, with little to no effect on the starvation response. Alfy is recruited to intracellular inclusions and scaffolds a complex between p62(SQSTM1)-positive proteins and the autophagic effectors Atg5, Atg12, Atg16L and LC3. Alfy overexpression leads to elimination of aggregates in an Atg5-dependent manner, and likewise, to protection in a neuronal and Drosophila model of polyglutamine toxicity. We propose that Alfy plays a key role in selective macroautophagy, by bridging cargo to the molecular machinery that builds autophagosomes.

Genetic Parkinson disease (PD) has been associated with mutations in PINK1, a gene encoding a mitochondrial kinase implicated in the regulation of mitochondrial degradation. While the studies so far examined PINK1 function in non-neuronal systems or through PINK1 knockdown approaches, there is an imperative to examine the role of endogenous PINK1 in appropriate human-derived and biologically relevant cell models. Here we report the generation of induced pluripotent stem (iPS) cells from skin fibroblasts taken from three PD patients with nonsense (c.1366C>T; p.Q456X) or missense mutations (c.509T>G; p.V170G) in the PINK1 gene. These cells were differentiated into dopaminergic neurons that upon mitochondrial depolarization showed impaired recruitment of lentivirally expressed Parkin to mitochondria, increased mitochondrial copy number and upregulation of PGC-1α, an important regulator of mitochondrial biogenesis. Importantly, these alterations were corrected by lentiviral expression of wild-type PINK1 in mutant iPS cell-derived PINK1 neurons. In conclusion, our studies suggest that fibroblasts from genetic PD can be reprogrammed and differentiated into neurons. These neurons exhibit distinct phenotypes that should be amenable to further mechanistic studies in this relevant biological context.

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This study examined the difficulties of running online physical education classes in the context of coronavirus disease 2019 (COVID-19) and used the findings to develop an efficient operation plan to address these difficulties. Six middle and high school physical education teachers participated; three were experts in online physical education and active in the Korea Council School Physical Education Promotion, and three were recommended teachers making efforts to improve the online classes offered by the Korea Ministry of Education. A qualitative case study method employing phenomenological procedures to collect and analyze the data was used. The difficulties of operating middle and high school online physical education classes for the first time included (1) the monotony of the classes within their limited environmental conditions and limited educational content that did not adequately convey the value of physical education, (2) trial-and-error methods applied nationwide, resulting from a lack of expertise in operating online physical education classes, and (3) very limited evaluation guidelines proposed by the Korea Ministry of Education, which made systematic evaluation with online methods impossible. To address the identified problems and facilitate the efficient operation of online physical education classes, changes in strategic learning methods are needed to understand online physical education characteristics and thereby better communicate the value of physical education. It is also necessary to cultivate teaching expertise through sharing online physical education classes, where collaboration among physical education teachers is central. In addition, evaluation processes should be less formal to encourage active student participation.