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Aim: A keloid is a fibroproliferative cutaneous disorder secondary to skin injury, caused by an imbalance in fibroblast proliferation and apoptosis. However, the pathogenesis is not fully understood. In this study, candidate genes for keloid were identified and used to construct a diagnostic model. Methods: Three datasets related to keloids were downloaded from NCBI Gene Expression Omnibus. Fibroblast-related genes were screened, and fibroblast scores for the samples were determined. Then, a weighted gene co-expression network analysis (WGCNA) was used to identify modules and genes associated with keloids and the fibroblast score. Differentially expressed genes (DEGs) between keloid and control samples were identified and compared with fibroblast-related genes and genes in the modules. Overlapping genes were evaluated using functional enrichment analyses. Signature genes were further screened, and a diagnostic model was constructed. Finally, correlations between immune cell frequences and signature genes were analyzed. Results: In total, 124 fibroblast-related genes were obtained, and the fibroblast score was an effective indicator of the sample type. WGCNA revealed five modules that were significantly correlated with both the disease state and fibroblast scores, including 1760 genes. Additionally, 589 DEGs were identified, including 16 that overlapped with fibroblast-related genes and genes identified in the WGCNA. These genes were related to cell proliferation and apoptosis and were involved in FoxO, Rap1, p53, Ras, MAPK, and PI3K-Akt pathways. Finally, a six fibroblast-related gene signature (CCNB1, EGFR, E2F8, BTG1, TP63, and IGF1) was identified and used for diagnostic model construction. The proportions of regulatory T cells and macrophages were significantly higher in keloid tissues than in controls. Conclusion: The established model based on CCNB1, EGFR, E2F8, BTG1, TP63, and IGF1 showed good performance and may be useful for keloid diagnosis.

Overaggressive liposuction of the infragluteal region can lead to iatrogenic infragluteal fold deformity and result in esthetic defects in the gluteal contour. We report a case of using autologous fat transplantation to correct severe post-liposuction infragluteal fold deformity. In the process of reconstruction, the patient experienced fat graft overabsorption, fat graft translocation, and gluteal ptosis aggravation. Despite multiple operations, the effect of fat transplantation was limited. In conclusion, severe post-liposuction infragluteal deformity is very difficult to correct. The infragluteal region should be preserved during liposuction to avoid deformity.

Target class-focused drug discovery has a strong track record in pharmaceutical research, yet public domain data indicate that many members of protein families remain unliganded. Here we present a systematic approach to scale up the discovery and characterization of small molecule ligands for the WD40 repeat (WDR) protein family. We developed a comprehensive suite of protocols for protein production, crystallography, and biophysical, biochemical, and cellular assays. A pilot hit-finding campaign using DNA-encoded chemical library selection followed by machine learning (DEL-ML) to predict ligands from virtual libraries yielded first-in-class, drug-like ligands for 7 of the 16 WDR domains screened, thus demonstrating the broader ligandability of WDRs. This study establishes a template for evaluation of protein family wide ligandability and provides an extensive resource of WDR protein biochemical and chemical tools, knowledge, and protocols to discover potential therapeutics for this highly disease-relevant, but underexplored target class.

Bfl-1 is overexpressed in both hematological and solid tumors; therefore, inhibitors of Bfl-1 are highly desirable. A DNA-encoded chemical library (DEL) screen against Bfl-1 identified the first known reversible covalent small-molecule ligand for Bfl-1. The binding was validated through biophysical and biochemical techniques, which confirmed the reversible covalent mechanism of action and pointed to binding through Cys55. This represented the first identification of a cyano-acrylamide reversible covalent compound from a DEL screen and highlights further opportunities for covalent drug discovery through DEL screening. A 10-fold improvement in potency was achieved through a systematic SAR exploration of the hit. The more potent analogue compound 13 was successfully cocrystallized in Bfl-1, revealing the binding mode and providing further evidence of a covalent interaction with Cys55., Competing Interests: The authors declare no competing financial interest., (© 2024 American Chemical Society.)

DNA-encoded chemical library (DEL) technology provides a time- and cost-efficient method to simultaneously screen billions of compounds for their affinity to a protein target of interest. Here we report its use to identify a novel chemical series of inhibitors of the thioesterase activity of polyketide synthase 13 (Pks13) from Mycobacterium tuberculosis (Mtb). We present three chemically distinct series of inhibitors along with their enzymatic and Mtb whole cell potency, the measure of on-target activity in cells, and the crystal structures of inhibitor-enzyme complexes illuminating their interactions with the active site of the enzyme. One of these inhibitors showed a favorable pharmacokinetic profile and demonstrated efficacy in an acute mouse model of tuberculosis (TB) infection. These findings and assay developments will aid in the advancement of TB drug discovery.

Dysregulation of IL17A drives numerous inflammatory and autoimmune disorders with inhibition of IL17A using antibodies proven as an effective treatment. Oral anti-IL17 therapies are an attractive alternative option, and several preclinical small molecule IL17 inhibitors have previously been described. Herein, we report the discovery of a novel class of small molecule IL17A inhibitors, identified via a DNA-encoded chemical library screen, and their subsequent optimization to provide in vivo efficacious inhibitors. These new protein-protein interaction (PPI) inhibitors bind in a previously undescribed mode in the IL17A protein with two copies binding symmetrically to the central cavities of the IL17A homodimer.

Evasion of apoptosis is critical for the development and growth of tumors. The pro-survival protein myeloid cell leukemia 1 (Mcl-1) is an antiapoptotic member of the Bcl-2 family, associated with tumor aggressiveness, poor survival, and drug resistance. Development of Mcl-1 inhibitors implies blocking of protein-protein interactions, generally requiring a lengthy optimization process of large, complex molecules. Herein, we describe the use of DNA-encoded chemical library synthesis and screening to directly generate complex, yet conformationally privileged macrocyclic hits that serve as Mcl-1 inhibitors. By applying a conceptual combination of conformational analysis and structure-based design in combination with a robust synthetic platform allowing rapid analoging, we optimized in vitro potency of a lead series into the low nanomolar regime. Additionally, we demonstrate fine-tuning of the physicochemical properties of the macrocyclic compounds, resulting in the identification of lead candidates 57 / 59 with a balanced profile, which are suitable for future development toward therapeutic use.

WD40 repeat-containing protein 91 (WDR91) regulates early-to-late endosome conversion and plays vital roles in endosome fusion, recycling, and transport. WDR91 was recently identified as a potential host factor for viral infection. We employed DNA-encoded chemical library (DEL) selection against the WDR domain of WDR91, followed by machine learning to predict ligands from the synthetically accessible Enamine REAL database. Screening of predicted compounds identified a WDR91 selective compound 1 , with a K D of 6 ± 2 μM by surface plasmon resonance. The co-crystal structure confirmed the binding of 1 to the WDR91 side pocket, in proximity to cysteine 487, which led to the discovery of covalent analogues 18 and 19 . The covalent adduct formation for 18 and 19 was confirmed by intact mass liquid chromatography-mass spectrometry. The discovery of 1 , 18 , and 19 , accompanying structure-activity relationship, and the co-crystal structures provide valuable insights for designing potent and selective chemical tools against WDR91 to evaluate its therapeutic potential.

DCAF1 is a substrate receptor of two distinct E3 ligases (CRL4 DCAF1 and EDVP), plays a critical physiological role in protein degradation, and is considered a drug target for various cancers. Antagonists of DCAF1 could be used toward the development of therapeutics for cancers and viral treatments. We used the WDR domain of DCAF1 to screen a 114-billion-compound DNA encoded library (DEL) and identified candidate compounds using similarity search and machine learning. This led to the discovery of a compound (Z1391232269) with an SPR K D of 11 μM. Structure-guided hit optimization led to the discovery of OICR-8268 ( 26e ) with an SPR K D of 38 nM and cellular target engagement with EC 50 of 10 μM as measured by cellular thermal shift assay (CETSA). OICR-8268 is an excellent tool compound to enable the development of next-generation DCAF1 ligands toward cancer therapeutics, further investigation of DCAF1 functions in cells, and the development of DCAF1-based PROTACs.