VAT has a significant association with cerebral small vessel disease, which was defined as WMH or lacunar infarct. Visceral obesity can be a potential therapeutic target for the prevention of cerebral small vessel disease.

The peptidyl-prolyl cis-trans isomerase, Pin1, acts as a unified signaling hub that is exploited in cancer to activate oncogenes and inactivate tumor suppressors, in particular through up-regulation of c-Myc target genes. However, despite considerable efforts, Pin1 has remained an elusive drug target. Here, we screened an electrophilic fragment library to discover covalent inhibitors targeting Pin1's active site nucleophile -Cys113, leading to the development of Sulfopin, a double-digit nanomolar Pin1 inhibitor. Sulfopin is highly selective for Pin1, as validated by two independent chemoproteomics methods, achieves potent cellular and in vivo target engagement, and phenocopies genetic knockout of Pin1. Although Pin1 inhibition had a modest effect on viability in cancer cell cultures, Sulfopin induced downregulation of c-Myc target genes and reduced tumor initiation and tumor progression in murine and zebrafish models of MYCN-driven neuroblastoma. Our results suggest that Sulfopin is a suitable chemical probe for assessing Pin1dependent pharmacology in cells and in vivo. Moreover, these studies indicate that Pin1 should be further investigated as a potential cancer target.

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20Complex neural circuitry requires stable connections formed by lengthy axons. To maintain these 21 functional circuits, fast transport delivers RNAs to distal axons where they undergo local translation. 22However, the mechanism that enables long distance transport of non-membrane enclosed organelles 23 such as RNA granules is not known. Here we demonstrate that a complex containing RNA and the RNA-24 binding protein (RBP) SFPQ interacts directly with a tetrameric kinesin containing the adaptor KLC1 and 25 the motor KIF5A. We show that binding of SFPQ to KIF5A/KLC1 motor complex is required for axon survival 26 and is impacted by KIF5A mutations that cause Charcot-Marie-Tooth (CMT) Disease. Moreover, 27 therapeutic approaches that bypass the need for local translation of SFPQ-bound proteins prevent axon 28 degeneration in CMT models. Collectively, these observations show that non-membrane enclosed 29 organelles can move autonomously and that replacing axonally translated proteins provides a therapeutic 30 approach to axonal degenerative disorders. 31 Results 75SFPQ granule, a non-membrane enclosed organelle, undergoes fast axonal transport. 77The RBP SFPQ is found in both cell bodies and axons of sensory neurons. However, the 78 mechanisms by which SFPQ and its critical RNA cargos are transported between these two locations is 79 not known. We utilized live cell imaging of DRG sensory neurons expressing Halo-tagged SFPQ to directly 80 visualize transport dynamics (Video 1). Fluorescent signal was enriched in the nucleus and was also 81 evident as discrete granules in the soma and axons, a pattern similar to the distribution of endogenous 82 SFPQ (Cosker et al., 2016). Consistent with the presence of intrinsically disordered regions within the 83 SFPQ coding sequence, Halo-tagged SFPQ granules exhibited liquid like properties during time-lapse 84 imaging (Gopal, Nirschl, Klinman, & Holzbaur, 2017), as the size and shape of SFPQ granules remained 85 constant at approximately 1 µm in diameter during the stationary phase, but the granules expanded and 86 elongated as they move ( Figure 1A and 1B). The majority of the Halo-tagged SFPQ granules in axons 87 were motile, either moving by retrograde transport (~48%), or anterograde transport (~28%), with the 88 remainder in stationary phase (~25%) ( Figure 1C and Figure 1-figure supplement 1A). SFPQ granules 89 exhibit an average anterograde velocity of 0.89 ± 0.08 μm/sec and average anterograde cumulative 90 displacement of 21.02 ± 2.49 μm, with an average retrograde velocity of 0.80 ± 0.04 μm/sec retrograde 91 and average retrograde cumulative displacement of 32.02 ± 2.45 μm (Figure 1D, Figure 1-figure 92 supplement 1B-E). Together, the velocity and the characteristics of movement indicate that the SFPQ-93 granules are non-membrane enclosed organelles that move in both directions by microtubule-94 dependent fast axonal transport, using a kinesin motor for anterograde and the more highly processive95 dynein motor for retrograde movements.96 97 98 99 5 SFPQ preferentiall...

Robust and efficient enzymes are essential modules for metabolic engineering and synthetic biology strategies across biological systems to engineer whole-cell biocatalysts. By condensing an acyl-CoA and an alcohol, alcohol acyltransferases (AATs) can serve as an interchangeable metabolic module for microbial biosynthesis of a diverse class of ester molecules with broad applications as flavors, fragrances, solvents, and drop-in biofuels. However, the current lack of robust and efficient AATs significantly limits their compatibility with heterologous precursor pathways and microbial hosts. Through bioprospecting and rational protein engineering, we identified and repurposed chloramphenicol acetyltransferases (CATs) from mesophilic prokaryotes to function as robust and efficient AATs compatible with at least 21 alcohol and 8 acyl-CoA substrates for microbial biosynthesis of linear, branched, saturated, unsaturated and/or aromatic esters. By plugging the best engineered CAT (CATec3 Y20F) into the gram-negative mesophilic bacterium Escherichia coli, we demonstrated that the recombinant strain could effectively convert various alcohols into desirable esters, for instance, achieving a titer of 13.9 g/L isoamyl acetate with 95% conversion by fed-batch fermentation. The recombinant E. coli was also capable of simulating the ester profile of roses with high conversion (> 97%) and titer (> 1 g/L) from fermentable sugars at 37oC. Likewise, a recombinant gram-positive, cellulolytic, thermophilic bacterium Clostridium thermocellum harboring CATec3 Y20F could produce many of these esters from recalcitrant cellulosic biomass at elevated temperatures (>50oC) due to the engineered enzyme' s remarkable thermostability. Overall, the engineered CATs can serve as a robust and efficient platform for designer ester biosynthesis from renewable and sustainable feedstocks.