ABSTRACT
Charcot-Marie-Tooth disease type 1A (CMT1A) is the most common inherited peripheral neuropathy caused by a 1.5 megabase tandem duplication of chromosome 17 harboring the PMP22 gene. This dose-dependent overexpression of PMP22 results in disrupted Schwann cell myelination of peripheral nerves. To get better insights into the underlying pathogenic mechanisms in CMT1A, we investigated the role of PMP22 duplication on cellular homeostasis in CMT1A mouse models and in patient-derived induced pluripotent stem cells differentiated into Schwann cell precursors (iPSC-SCPs). We performed lipidomic profiling and bulk RNA sequencing on sciatic nerves of two developing CMT1A mouse models and on CMT1A patient derived iPSC-SCPs. For the sciatic nerves of the CMT1A mice, cholesterol and lipid metabolism was dose-dependently downregulated throughout development. For the CMT1A iPSC-SCPs, transcriptional analysis unveiled a strong suppression of genes related to autophagy and lipid metabolism. Gene ontology enrichment analysis identified disturbances in pathways related to plasma membrane components and cell receptor signaling. Lipidomic analysis confirmed the severe dysregulation in plasma membrane lipids, particularly sphingolipids, in CMT1A iPSC-SCPs. Furthermore, we identified reduced lipid raft dynamics, disturbed plasma membrane fluidity, and impaired cholesterol incorporation and storage, all of which could result from altered lipid storage homeostasis in the patient-derived CMT1A iPSC-SCPs. Importantly, this phenotype could be rescued by stimulating autophagy and lipolysis. We conclude that PMP22 duplication disturbs intracellular lipid storage and leads to a more disordered plasma membrane due to an alteration in the lipid composition, which ultimately may lead to impaired axo-glial interactions. Moreover, targeting lipid handling and metabolism could hold promise for the treatment of CMT1A patients.
ABSTRACT
OBJECTIVE: Epidemiological studies show that shift workers are at increased risk for type 2 diabetes. As modern societies increasingly require shift work, it seems crucial to determine whether there are long-lasting health effects of rotational shifts. METHODS: This study examined the after-effects of 4 weeks of time-restricted feeding (TRF) during the light period (= light-fed) in rats, an animal model for shift work. This study also included a TRF-dark (= dark-fed) control group. The aligned and misaligned feeding times of light and dark feeding are associated with poor and good health outcomes, respectively. Several physiological measures were monitored continuously; blood, liver, brown adipose tissue, and soleus and gastrocnemius muscle were collected following 11 days of ad libitum (AL) feeding after ending the TRF. RESULTS: In the dark-fed animals, the day/night differences in food intake, activity, and respiratory exchange ratio were still enhanced at the end of the experiment. Light-fed animals displayed the smallest day/night differences for these measures, as well as for body temperature. In both the light- and dark-fed animals, rhythms in plasma glucose, nonesterified fatty acids, and gene expression had not fully recovered after 11 days of AL feeding. Importantly, the effects on gene expression were both tissue and gene dependent. CONCLUSIONS: Our data indicate that rotational shift workers may have an increased risk of long-lasting disturbed rhythms in several physiological measures after a period of shift work. Clearly, such disturbances may harm their health.
