Backbone N-methylation of peptides: Advances in synthesis and applications in pharmaceutical drug development.

Peptide-based drugs have garnered considerable attention in recent years owing to their increasingly crucial role in the treatment of diverse diseases. However, the limited pharmacokinetic properties of peptides have hindered their full potential. One prominent strategy for enhancing the druggability of peptides is N-methylation, which involves the addition of a methyl group to the nitrogen atom of the peptide backbone. This modification significantly improves the stability, bioavailability, receptor binding affinity and selectivity of peptide drug candidates. In this review, we provide a comprehensive overview of the advancements in synthetic methods for N-methylated peptide synthesis, as well as the associated limitations. Moreover, we explore the versatile effects of N-methylation on various aspects of peptide properties. Furthermore, we emphasize the efforts dedicated to N-methylated peptide pharmaceuticals that have successfully obtained marketing approval.

Climatology of Mesosphere and Lower Thermosphere Residual Circulations and Mesopause Height Derived From SABER Observations.

In the mesosphere and lower thermosphere (MLT) region, residual circulations driven by gravity wave breaking and dissipation significantly impact constituent distribution and the height and temperature of the mesopause. The distribution of CO2 can be used as a proxy for the residual circulations. Sounding of the Atmosphere using Broadband Emission Radiometry (SABER) CO2 volume mixing ratio (VMR) and temperature measurements from 2003 to 2020 are used to study the monthly climatology of MLT residual circulations and the mesopause height. Our analyses show that (a) mesopause height strongly correlates with the CO2 VMR vertical gradient during solstices; (b) mesopause height has a discontinuity at midlatitude in the summer hemisphere, with a lower mesopause height at mid-to-high latitudes as a result of adiabatic cooling driven by strong adiabatic upwelling; (c) the residual circulations have strong seasonal variations at mid-to-high latitudes, but they are more uniform at low latitudes; and (d) the interannual variability of the residual circulations and mesopause height is larger in the Southern Hemisphere (SH; 4-5 km) than in the Northern Hemisphere (NH; 0.5-1 km).

Empagliflozin-Enhanced Antioxidant Defense Attenuates Lipotoxicity and Protects Hepatocytes by Promoting FoxO3a- and Nrf2-Mediated Nuclear Translocation via the CAMKK2/AMPK Pathway.

Lipotoxicity is an important factor in the development and progression of nonalcoholic steatohepatitis. Excessive accumulation of saturated fatty acids can increase the substrates of the mitochondrial electron transport chain in hepatocytes and cause the generation of reactive oxygen species, resulting in oxidative stress, mitochondrial dysfunction, loss of mitochondrial membrane potential, impaired triphosphate (ATP) production, and fracture and fragmentation of mitochondria, which ultimately leads to hepatocellular inflammatory injuries, apoptosis, and necrosis. In this study, we systematically investigated the effects and molecular mechanisms of empagliflozin on lipotoxicity in palmitic acid-treated LO2 cell lines. We found that empagliflozin protected hepatocytes and inhibited palmitic acid-induced lipotoxicity by reducing oxidative stress, improving mitochondrial functions, and attenuating apoptosis and inflammation responses. The mechanistic study indicated that empagliflozin significantly activated adenosine 5'-monophosphate (AMP)-activated protein kinase alpha (AMPKα) through Calcium/Calmodulin dependent protein kinase kinase beta (CAMKK2) instead of liver kinase B1 (LKB1) or TGF-beta activated kinase (TAK1). The activation of empagliflozin on AMPKα not only promoted FoxO3a phosphorylation and thus forkhead box O 3a (FoxO3a) nuclear translocation, but also promoted Nrf2 nuclear translocation. Furthermore, empagliflozin significantly upregulated the expressions of antioxidant enzymes superoxide dismutase (SOD) and HO-1. In addition, empagliflozin did not attenuate lipid accumulation at all. These results indicated that empagliflozin mitigated lipotoxicity in saturated fatty acid-induced hepatocytes, likely by promoting antioxidant defense instead of attenuating lipid accumulation through enhanced FoxO3a and Nrf2 nuclear translocation dependent on the CAMKK2/AMPKα pathway. The CAMKK2/AMPKα pathway might serve as a promising target in treatment of lipotoxicity in nonalcoholic steatohepatitis.

RNA-binding Protein MBNL2 regulates Cancer Cell Metastasis through MiR-182-MBNL2-AKT Pathway.

The aberrant expression of RNA-binding proteins (RBPs) plays important roles in the occurrence and progression of cancer. MBNL2 is a member of the RNA binding protein MBNL family that is widely expressed in mammalian cells. We report here that MBNL2 is downregulated in breast, lung and liver cancer tissues, the promoter methylation levels of MBNL2 are higher in cancer tissues than normal tissues. The enrichment analysis of MBNL2 correlated genes indicates the potential function of MBNL2 on cancer progression. MBNL2 regulates cancer cell migration and invasion by modulating PI3K/AKT-mediated epithelial-mesenchymal transition. PI3K/AKT inhibitor overcomes the promotive effect of shMBNL2 on metastasis. The expression of MBNL2 is directly targeted by miR-182. miR-182 is upregulated in breast, lung and liver cancers and has good potential for cancer diagnosis. miR-182 promotes cancer cell migration and invasion by inhibiting the expression of MBNL2. Re-introduction of exogenous MBNL2 reverses the promotive effect of miR-182 on metastasis. Collectively, these findings suggest that MBNL2 plays a tumor suppressive function through miR-182-MBNL2-AKT-EMT signaling pathways.

USP30-AS1 contributes to mitochondrial quality control in glioblastoma cells.

Glioblastoma is the most serious type of brain cancer with poor prognosis. Here, using the publicly available glioma database, we identified that USP30-AS1, an antisense lncRNA locating on the opposite strand of USP30 locus, is upregulated in human gliomas, particularly in high grade glioma. High level of USP30-AS1 is correlated with poor survival in both primary and recurrent glioma patients. USP30-AS1 regulates mitochondrial homeostasis and mitophagy in glioblastoma cells. Knockdown of USP30-AS1 decreases mitochondrial protein expression and mitochondrial mass, promotes mitochondrial uncoupler-induced mitophagy. However, USP30-AS1 does not regulate USP30 expression in a cis-regulatory manner. In summary, this study proposed that USP30-AS1 may serve as a valuable prognostic marker for gliomas. USP3-AS1 is a negative regulator of mitophagy and the regulatory effect is USP30-independent. USP30-AS1 mediated repression of mitophagy may contribute to the loss of mitochondrial homeostasis and tumor development in glioma.

Transport of Nitric Oxide Via Lagrangian Coherent Structures Into the Top of the Polar Vortex.

The energetic particle precipitation (EPP) indirect effect (IE) refers to the downward transport of reactive odd nitrogen (NOx = NO + NO2) produced by EPP (EPP-NOx) from the polar winter mesosphere and lower thermosphere to the stratosphere where it can destroy ozone. Previous studies of the EPP IE examined NOx descent averaged over the polar region, but the work presented here considers longitudinal variations. We report that the January 2009 split Arctic vortex in the stratosphere left an imprint on the distribution of NO near the mesopause, and that the magnitude of EPP-NOx descent in the upper mesosphere depends strongly on the planetary wave (PW) phase. We focus on an 11-day case study in late January immediately following the 2009 sudden stratospheric warming during which regional-scale Lagrangian coherent structures (LCSs) formed atop the strengthening mesospheric vortex. The LCSs emerged over the north Atlantic in the vicinity of the trough of a 10-day westward traveling planetary wave. Over the next week, the LCSs acted to confine NO-rich air to polar latitudes, effectively prolonging its lifetime as it descended into the top of the polar vortex. Both a whole atmosphere data assimilation model and satellite observations show that the PW trough remained coincident in space and time with the NO-rich air as both migrated westward over the Canadian Arctic. Estimates of descent rates indicate five times stronger descent inside the PW trough compared to other longitudes. This case serves to set the stage for future climatological analysis of NO transport via LCSs.

MBNL2 Regulates DNA Damage Response via Stabilizing p21.

RNA-binding proteins are frequently dysregulated in human cancer and able to modulate tumor cell proliferation as well as tumor metastasis through post-transcriptional regulation on target genes. Abnormal DNA damage response and repair mechanism are closely related to genome instability and cell transformation. Here, we explore the function of the RNA-binding protein muscleblind-like splicing regulator 2 (MBNL2) on tumor cell proliferation and DNA damage response. Transcriptome and gene expression analysis show that the PI3K/AKT pathway is enriched in MBNL2-depleted cells, and the expression of cyclin-dependent kinase inhibitor 1A (p21CDKN1A) is significantly affected after MBNL2 depletion. MBNL2 modulates the mRNA and protein levels of p21, which is independent of its canonical transcription factor p53. Moreover, depletion of MBNL2 increases the phosphorylation levels of checkpoint kinase 1 (Chk1) serine 345 (S345) and DNA damage response, and the effect of MBNL2 on DNA damage response is p21-dependent. MBNL2 would further alter tumor cell fate after DNA damage, MBNL2 knockdown inhibiting DNA damage repair and DNA damage-induced senescence, but promoting DNA damage-induced apoptosis.

Lower Thermospheric Material Transport via Lagrangian Coherent Structures.

We show that inter-model variation due to under-constraint by observations impacts the ability to predict material transport in the lower thermosphere. Lagrangian coherent structures (LCSs), indicating regions of maximal separation (or convergence) in a time-varying flow, are derived in the lower thermosphere from models for several space shuttle water vapor plume events. We find that inter-model differences in thermospheric transport manifest in LCSs in a way that is more stringent than mean wind analyses. LCSs defined using horizontal flow fields from the Specified Dynamics version of the Whole Atmosphere Community Climate Model with thermosphere-ionosphere eXtension (SD-WACCMX) at 109 km altitude are compared to Global Ultraviolet Imager (GUVI) observations of the space shuttle main engine plume. In one case, SD-WACCMX predicts an LCS ridge to produce spreading not found in the observations. LCSs and tracer transport from SD-WACCMX and from data assimilative WACCMX (WACCMX + DART) are compared to each other and to GUVI observations. Differences in the modeled LCSs and tracer positions appear between SD-WACCMX and WACCMX + DART despite the similarity of mean winds. WACCMX + DART produces better tracer transport results for a July 2006 event, but it is unclear which model performs better in terms of LCS ridges. For a February 2010 event, when mean winds differ by up to 50 m/s between the models, differences in LCSs and tracer trajectories are even more severe. Low-pass filtering the winds up to zonal wavenumber 6 reduces but does not eliminate inter-model LCS differences. Inter-model alignment of LCSs improves at a lower 60 km altitude.