It is widely known that the dysregulation of non-coding RNAs (ncRNAs) and dysbiosis of the gut microbiome play significant roles in host development and the progression of various diseases. Emerging evidence has highlighted the bidirectional interplay between ncRNAs and the gut microbiome. This article aims to review the current understanding of the molecular mechanisms underlying the crosstalk between ncRNAs, especially microRNA (miRNA), and the gut microbiome in the context of development and diseases, such as colorectal cancer, inflammatory bowel diseases, neurological disorders, obesity, and cardiovascular disease. Ultimately, this review seeks to provide a foundation for exploring the potential roles of ncRNAs and gut microbiome interactions as biomarkers and therapeutic targets for clinical diagnosis and treatment, such as ncRNA mimics, antisense oligonucleotides, and small-molecule compounds, as well as probiotics, prebiotics, and diets.

O-GlcNAcase (OGA) is the sole eraser for the intracellular O-linked N-acetylglucosamine (O-GlcNAc). OGA has many roles in distinct biological processes, such as cancer and embryonic stem cells, but its precise regulatory mechanism is far from being understood. Herein we studied the small ubiquitin-like modifier (SUMO) modification of OGA, and found that OGA is SUMOylated at K358. SUMOylation targets OGA to the chaperone-mediated autophagy (CMA) pathway, which shunts client proteins to the lysosome for degradation. We demonstrate that SUMOylation increases the association between OGA and the heat shock cognate protein 70 (HSC70), the CMA chaperone, and facilitates OGA further degradation. We further mapped a SUMO-interacting motif (SIM) (VLIFD, aa. 195-199) on HSC70. Notably, HSC70-SIM is essential for affinity with other CMA client proteins, such as PKM2. We thus posit that the SIM of HSC70 binds SUMOylated client proteins in a lock-and-key manner to confer substrate selectivity during CMA. To further test our hypothesis, we used label-free quantitative mass spectrometry to study the HSC70-SIM mutant interactome, and generated a proteome-wide SUMO-mediated CMA client pool. We then validated this model by studying YEATS domain containing 2 (YEATS2) from the protein pool, and demonstrated that YEATS2 is SUMOylated at K592, targeting it to CMA. Our work uncovers the SUMO-SIM interaction as a fundamental mechanism governing CMA substrate selectivity and identifies a potential CMA client proteome to deepen our understanding of its pathophysiological relevance.

Emerging evidence has illuminated the critical role of gut microbiota in regulating health and longevity. In this review, we systematically summarize the changes in gut microbiota associated with aging and age-related diseases, including gastrointestinal tract diseases, neurodegenerative disorders, and metabolic dysfunction syndromes. We will also explore the underlying mechanisms, particularly those related to metabolic and immune pathways, highlighting a feedback loop model of "microbial dysbiosis-metabolic dysfunction-chronic inflammation". Furthermore, current interventions related to gut microbiota, including dietary strategies, fecal microbiota transplantation, prebiotics, probiotics, and phage therapies, will be discussed. We will also explore cutting-edge technologies such as artificial intelligence (AI)-driven microbiome analysis, microbial engineering, and organ-on-chip systems, as well as the challenges that remain in this field. Overall, this review provides a theoretical framework and directional guidance for rejuvenating aging and age-related diseases via targeting the gut microbiota.

O-linked N-acetylglucosamine (O-GlcNAc) is the most abundant mono-saccharide modification occurring in the cytoplasm, nucleus, and mitochondria. The recent advent of mass spectrometry technology has enabled the identification of abundant O-GlcNAc transferase (OGT) substrates in diverse biological processes, such as cell cycle progression, replication, and DNA damage response. Herein we report the O-GlcNAcylation of Replication Protein A2 (RPA2), a component of the heterotrimeric RPA complex pivotal for DNA metabolism. We found that RPA2 interacts with OGT, and a topoisomerase II inhibitor, etoposide, diminishes the association. Using higher-energy collisional dissociation mass spectrometry, we mapped RPA2 O-GlcNAc sites to be Ser-4/Ser-8, which are well-known PIKK-dependent RPA2 phosphorylation sites involved in checkpoint activation upon replication stress. We further demonstrated that Ser-4/Ser-8 O-GlcNAcylation antagonizes phosphorylation and impairs downstream Chk1 activation. Moreover, RPA2 O-GlcNAcylation sustains H2AX phosphorylation upon etoposide treatment and promotes inappropriate cell cycle progression, indicative of checkpoint defects. Our work not only unveils a new OGT substrate, but also underscores the distinct roles of OGT in replication versus replication stress.