Transcriptional regulatory networks of tumor-associated macrophages that drive malignancy in mesenchymal glioblastoma.

BACKGROUND: Glioblastoma (GBM) is a complex disease with extensive molecular and transcriptional heterogeneity. GBM can be subcategorized into four distinct subtypes; tumors that shift towards the mesenchymal phenotype upon recurrence are generally associated with treatment resistance, unfavorable prognosis, and the infiltration of pro-tumorigenic macrophages. RESULTS: We explore the transcriptional regulatory networks of mesenchymal-associated tumor-associated macrophages (MA-TAMs), which drive the malignant phenotypic state of GBM, and identify macrophage receptor with collagenous structure (MARCO) as the most highly differentially expressed gene. MARCO CONCLUSIONS: Collectively, our study characterizes the global transcriptional profile of TAMs driving mesenchymal GBM pathogenesis, providing potential therapeutic targets for improving the effectiveness of GBM immunotherapy

Enhanced recovery after vascular surgery

INTRODUCTION: A multimodal perioperative care measure, the enhanced recovery after surgery (ERAS) method, is intended to accomplish early recovery following surgical procedures. It aims to preserve preoperative organ function and mitigate the significant stress response that typically occurs during recovery. EVIDENCE ACQUISITION: The goal of this systematic review and meta-analysis was to evaluate the advantages of enhanced recovery in the scope of vascular surgery. Following PRISMA Guidelines, a systematic search was conducted on various electronic reference databases (Web of Sciences, PubMed and Cochrane library). The keywords employed were (“Enhanced Recovery After Surgery” OR “ERAS”) AND (vascular) AND (“surgery” OR “operation” OR “procedure”). Inclusion criteria are articles published in English, and full-text was available, published between 2013-2023. Data was obtained on hospitalization duration, in hospital mortality, and post-surgical morbidity. EVIDENCE SYNTHESIS: Five hundred seventeen articles were identified and seven papers involving 1954 patients included for systematic review. The incidence of postoperative morbidity demonstrated a significant reduction when utilizing the ERAS approach in comparison to non-ERAS protocols (OR=0.21 [95%CI, 0.08 to 0.54], P=0.001). Additionally, the implementation of an ERAS protocol resulted in a notable reduction in hospitalization duration (MD=-0.59 [95%CI, -1.13 to -0.04], P=0.04). Furthermore, no significant difference was identified in hospital mortality rates. CONCLUSIONS: The multimodal perioperative care approach known as Enhanced Recovery After Surgery (ERAS) method is intended to facilitate immediate postoperative recovery by safeguarding preoperative organ function and reducing the visceral stress response post-surgery. The utilization of ERAS, coupled with enhanced perioperative care, brings substantial advantages to patients undergoing vascular surgery as well as those undergoing a range of other surgical specialties

Promising Therapeutic Efficacy of GC1118, an Anti-EGFR Antibody, against KRAS Mutation-Driven Colorectal Cancer Patient-Derived Xenografts

Epidermal growth factor receptor (EGFR)-targeted monoclonal antibodies, including cetuximab and panitumumab, are used to treat metastatic colorectal cancer (mCRC). However, this treatment is only effective for a small subset of mCRC patients positive for the wild-type KRAS GTPase. GC1118 is a novel, fully humanized anti-EGFR IgG1 antibody that displays potent inhibitory effects on high-affinity EGFR ligand-induced signaling and enhanced antibody-mediated cytotoxicity. In this study, using 51 CRC patient-derived xenografts (PDXs), we showed that KRAS mutants expressed remarkably elevated autocrine levels of high-affinity EGFR ligands compared with wild-type KRAS. In three KRAS-mutant CRCPDXs, GC1118 was more effective than cetuximab, whereas the two agents demonstrated comparable efficacy against three wild-type KRAS PDXs. Persistent phosphatidylinositol-3-kinase (PI3K)/AKT signaling was thought to underlie resistance to GC1118. In support of these findings, a preliminary improved anti-cancer response was observed in a CRC PDX harboring mutated KRAS with intrinsically high AKT activity using GC1118 combined with the dual PI3K/mammalian target of rapamycin (mTOR)/AKT inhibitor BEZ-235, without observed toxicity. Taken together, the superior antitumor efficacy of GC1118 alone or in combination with PI3K/mTOR/AKT inhibitors shows great therapeutic potential for the treatment of KRAS-mutant mCRC with elevated ratios of high- to low-affinity EGFR ligands and PI3K-AKT pathway activation

Correction: Functional implications of hexameric assembly of RraA proteins from Vibrio vulnificus.

[This corrects the article DOI: 10.1371/journal.pone.0190064.]

Functional implications of hexameric assembly of RraA proteins from Vibrio vulnificus.

RNase E has a pivotal role in the degradation and processing of RNAs in Escherichia coli, and protein inhibitors RraA and RraB control its enzymatic activity. The halophilic pathogenic bacterium Vibrio vulnificus also expresses orthologs of RNase E and RraA-RNase EV, RraAV1, and RraAV2 (herein renamed as VvRNase E, VvRraA1, and VvRraA2). A previous study showed that VvRraA1 actively inhibits the ribonucleolytic activity of VvRNase E by interacting with the C-terminal region of VvRNase E. However, the molecular mechanism underlying the effect of VvRraA1 on the ribonucleolytic activity of VvRNase E has not yet been elucidated. In this study, we report that the oligomer formation of VvRraA proteins affects binding efficiency to VvRNase E as well as inhibitory activity on VvRNase E action. The hexameric structure of VvRraA1 was converted to lower oligomeric forms when the Cys 9 residue was substituted with an Asp residue (VvRraA1-C9D), showing decreased inhibitory activity of VvRraA1 on VvRNase E in vivo. These results indicated that the intermolecular disulfide linkage contributed critically to the hexamerization of VvRraA1 for its proper function. On the contrary, the VvRraA2 that existed in a trimeric state did not bind to or inhibit VvRNase E. An in vitro cleavage assay further showed the reduced inhibitory effect of VvRraA-C9D on VvRNase E activity compared to wild-type VvRraA1. These findings provide insight into how VvRraA proteins can regulate VvRNase E action on its substrate RNA in V. vulnificus. In addition, based on structural and functional comparison of RraA homologs, we suggest that hexameric assembly of RraA homologs may well be required for their action on RNase E-like proteins

Oligomerization state of VvRraA proteins. Size exclusion chromatography with MALS of VvRraA proteins and their C9D mutants.

<p>The molecular sizes of the peaks were estimated by using MALS, as indicated. Based on these results, <i>a</i>, corresponds to 75.62 kDa (~hexamer); <i>b</i>, 16.3 kDa (monomer); <i>c</i>, 56.64 kDa (trimer); <i>d</i>, 19.68 kDa (monomer). Although the peak ‘<i>a</i>’ for the wild-type VvRraA1 is in between hexameric and trimeric sizes (~4.7 times higher than the monomeric mutant), it is most likely that VvRraA1 is a hexameric form since all the native RraA proteins are in the forms of a trimer or a hexamer.</p

Alignment of amino acid sequences of <i>E</i>. <i>coli</i> RraA and its orthologs in Gram-negative bacteria.

<p>(A) Alignment of amino acid sequence using CLUSTAL W. VvRraA1 and VvRraA2; Amino acid sequences of RraA homologs from <i>E</i>. <i>coli</i> (EcRraA), <i>Mycobacterium tuberculosis</i> (MtRraA), <i>P</i>. <i>aeruginosa</i> (PaRraA), <i>Vibrio cholerae</i> (VcRraA), <i>V</i>. <i>vulnificus</i> (VvRraA1 and VvRraA2) are used. Arrows indicate conserved Cys9 and Cys41 residues of RraA proteins. (B) A molecular model for the C9D mutant of <i>E</i>. <i>coli</i> RraA. The model of the mutant protein was built based on the wild-type structure of <i>E</i>. <i>coli</i> RraA (PDB code: 1Q5X). The subunits are displayed in different colors (cyan and yellow). The mutated Asp9 is positioned in the hydrophobic pocket lined with residues in gold at the interface between the two neighboring subunits, which would destabilize the oligomeric forms of the protein (left lower box). The near region of Cys9 structure is shown in the left upper box.</p

Inhibition of VvRraA1 and VvRraA1-C9D on the cleavage of p-BR10+hpT by VvRNase E <i>in vitro</i>.

<p>0.5 pmol of 5’-end-labeled p-BR10+hpT RNA was incubated with 1 pmol of VvRne with varying concentrations of VvRraA1 and VvRraA1-C9D, 50 pmol of VvRraA1, or 50 pmol of BSA in 20 μl of 1 × cleavage buffer at 37°C for 2 h for VvRne, VvRraA1 only, or BSA only controls. Samples were mixed with an equal volume of loading buffer, and then denatured at 65°C for 5 min and loaded onto a 12% polyacrylamide gel containing 8 M urea. The percentage of uncleaved p-BR10+hpT in the gel was quantitated using a phosphorimager and OptiQuant software.</p

Interactions of VvRNase E with VvRraA proteins.

<p>Hexahistidine-tagged VvRraA1, VvRraA1-C9D, VvRraA2, VvRraA2-C9D, and the GST-fused VvRne (612–816 residues) were expressed and purified as described in the Methods section. The GST-fused VvRne protein was bound to GSH resin and incubated with VvRraA proteins and their C9D mutant proteins. Then, the proteins were eluted and the fractions were analyzed using SDS-PAGE. The protein bands were stained with Coomassie blue. Only VvRraA1 could tightly bind to VvRne.</p