Wnt Genes Research Articles

Molecular mechanism of an aspirin compound for alleviation of osteoporosis using nerwork pharmacology and molecular docking

The aspirin compound (AC) is commonly found to have a wide range ofpharmacological activities. This study aimed to investigate the underlying mechanism ofthe anti-osteoporotic (anti-OP) activity of AC using network pharmacology and moleculardocking approaches. First, AC targets were identified using the GeneCards database, andsecond, OP-related targets were mined using a combination of the GeneCards andDisGeNet databases. The intersection targets from the Genecards, AC, and OP databaseswere considered candidate targets and were utilized to calculate protein-protein interactionsbetween targets. We discovered a C4A intersection target in the Genecards, AC, and OPdatabases. This is useful for molecular binding. In addition, we obtained 11 additionalprospective targets that may be used to attach the AC molecule to these targets. At theintersection region of the AC and OP target groups are the target genes HLA-DQA1, HLADQB1, RPL31, SATB2, SP1, and WNT1. And at the intersection region of the GeneCardsand OP databases, we discovered the anti-OP target genes CTSK, PDIA2, RARG, andTBC1D8. They are crucial binding targets that interact with other protein targets.According to the protein-protein interaction network, C4A showed the highest bindingcapacity with other proteins. The gene ontology (GO) analysis performed in this workfound the 10 biological processes, 10 cellular components, 9 molecular activities, and 13biological pathways. Furthermore, C4A and 10 candidate targets are associated withhomologous genes mainly involved in the signaling pathways of the Kyoto Encyclopediaof Genomics and Genomics (KEGG). The AC molecule was found to be highly bound to11 target candidates. This study identified candidate targets for AC to alleviate OP usingthe search for protein-protein interactions and the associated signaling pathways of thetargets to endorse AC against OP. This may help us understand the mechanism of action ofAC during OP treatment.

PANORAMA OF GASTRIC AND INTESTINAL CARCINOGENESIS: LITERATURE REVIEW

Purpose. The aim of the review is to analyze the modern data on the molecular mechanisms of stomach and intestinal carcinogenesis, which can be the basis for managing of malignant stomach and intestine neoplasms mortality risks’ reduction.
 Materials and methods. The review includes data from foreign and domestic studies published in the electronic bibliographic databases PubMed, Cyberleninka, Elibrary over the past 10 years.
 Results. It is given the analysis of Wnt and β-catenin gene family role in the pathogenesis of stomach and colon malignant tumors and the carcinogenic potential of Helicobacter pylori in gastric cancer, as well as the driver bacteria contributing to the development of colon cancer.
 Conclusion. Stomach cancer and colon cancer are heterogeneous diseases with diverse molecular and histological subtypes, having both similarities and differences in the patterns of cancer promotion. Progress in transparency genesis of these diseases is associated with an understanding of cytotoxic and genotoxic effects, oxidative stress and chronic inflammation mechanisms.

Wnt/β-Catenin Signaling Activation Induces Differentiation in Human Limbal Epithelial Stem Cells Cultured Ex Vivo.

Human limbal epithelial stem cells (hLESCs) continuously replenish lost or damaged human corneal epithelial cells. The percentage of stem/progenitor cells in autologous ex vivo expanded tissue is essential for the long-term success of transplantation in patients with limbal epithelial stem cell deficiency. However, the molecular processes governing the stemness and differentiation state of hLESCs remain uncertain. Therefore, we sought to explore the impact of canonical Wnt/β-catenin signaling activation on hLESCs by treating ex vivo expanded hLESC cultures with GSK-3 inhibitor LY2090314. Real-time qRT-PCR and microarray data reveal the downregulation of stemness (TP63), progenitor (SOX9), quiescence (CEBPD), and proliferation (MKI67, PCNA) genes and the upregulation of genes for differentiation (CX43, KRT3) in treated- compared to non-treated samples. The pathway activation was shown by AXIN2 upregulation and enhanced levels of accumulated β-catenin. Immunocytochemistry and Western blot confirmed the findings for most of the above-mentioned markers. The Wnt/β-catenin signaling profile demonstrated an upregulation of WNT1, WNT3, WNT5A, WNT6, and WNT11 gene expression and a downregulation for WNT7A and DKK1 in the treated samples. No significant differences were found for WNT2, WNT16B, WIF1, and DKK2 gene expression. Overall, our results demonstrate that activation of Wnt/β-catenin signaling in ex vivo expanded hLESCs governs the cells towards differentiation and reduces proliferation and stem cell maintenance capability.

Prohibitin Links Cell Cycle, Motility and Invasion in Prostate Cancer Cells.

Prohibitin (PHB) is a tumour suppressor gene with several different molecular activities. PHB overexpression leads to G1/S-phase cell cycle arrest, and PHB represses the androgen receptor (AR) in prostate cancer cells. PHB interacts with and represses members of the E2F family in a manner that may also be AR-linked, therefore making the AR:PHB:E2F interaction axis highly complex. PHB siRNA increased the growth and metastatic potential of LNCaP mouse xenografts in vivo. Conversely, PHB ectopic cDNA overexpression affected several hundred genes in LNCaP cells. Furthermore, gene ontology analysis showed that in addition to cell cycle regulation, several members of the WNT family were significantly downregulated (WNT7B, WNT9A and WNT10B), as well as pathways for cell adhesion. Online GEO data studies showed PHB expression to be decreased in clinical cases of metastatic prostate cancer, and to be correlated with higher WNT expression in metastasis. PHB overexpression reduced prostate cancer cell migration and motility in wound-healing assays, reduced cell invasion through a Matrigel layer and reduced cellular attachment. In LNCaP cells, WNT7B, WNT9A and WNT10B expression were also upregulated by androgen treatment and downregulated by androgen antagonism, indicating a role for AR in the control of these WNT genes. However, these WNTs were strongly cell cycle regulated. E2F1 cDNA ectopic expression and PHB siRNA (both cell cycle promoting effects) increased WNT7B, WNT9A and WNT10B expression, and these genes were also upregulated as cells were released from G1 to S phase synchronisation, indicating further cell cycle regulation. Therefore, the repressive effects of PHB may inhibit AR, E2F and WNT expression and its loss may increase metastatic potential in human prostate cancer.

Medical Care Use Among Patients with Monogenic Osteoporosis Due to Rare Variants in LRP5, PLS3, or WNT1

Pathogenic variants in the LRP5, PLS3, or WNT1 genes can significantly affect bone mineral density, causing monogenic osteoporosis. Much remains to be discovered about the phenotype and medical care needs of these patients. The purpose of this study was to examine the use of medical care among Dutch individuals identified between 2014 and 2021 with a pathogenic or suspicious rare variant in LRP5, PLS3, or WNT1. In addition, the aim was to compare their medical care utilization to both the overall Dutch population and the Dutch Osteogenesis Imperfecta (OI) population. The Amsterdam UMC Genome Database was used to match 92 patients with the Statistics Netherlands (CBS) cohort. Patients were categorized based on their harbored variants: LRP5, PLS3, or WNT1. Hospital admissions, outpatient visits, medication data, and diagnosis treatment combinations (DTCs) were compared between the variant groups and, when possible, to the total population and OI population. Compared to the total population, patients with an LRP5, PLS3, or WNT1 variant had 1.63 times more hospital admissions, 2.0 times more opened DTCs, and a greater proportion using medication. Compared to OI patients, they had 0.62 times fewer admissions. Dutch patients with an LRP5, PLS3, or WNT1 variant appear to require on average more medical care than the total population. As expected, they made higher use of care at the surgical and orthopedic departments. Additionally, they used more care at the audiological centers and the otorhinolaryngology (ENT) department, suggesting a higher risk of hearing-related problems.

Abstract A046: Integrated genomic analysis of primary prostate tumor foci and corresponding lymph node metastases identifies pathways associated with metastatic disease

Abstract Background: Prostate cancer (PCa) is a highly heterogeneous disease, and mortality is mainly due to metastases. However, the molecular underpinnings that lead to the initial steps of metastasis have not been well characterized. We have performed integrative whole exome sequencing and transcriptome analysis of primary prostate tumor foci and corresponding lymph node metastases (LNM). Design: Primary tumor foci (PTF) and LNM from 40 patients with high-risk PCa were analyzed by RNAseq. Two or more PTF and all available LNM greater than 0.4cm were subjected to sequencing. Of these 40 patients, 17 (42.5%) had LNM and 23 (57.5%) had benign LNs. A total of 155 tissue samples (97 PTF, 39 benign LNs, and 19 LNM) were sequenced and mapped to the human transcriptome with STAR mapper after QC trimming and removal of adapter sequences using TrimGalore. Differentially expressed genes between PTF, LNM, and benign LNs were identified using DESeq2, and gene set enrichment analysis was performed using WebGestalt. WES data was analyzed using GATK pipelines including Mutect2 and maftools. Results: A median of 57 million paired-end reads were obtained per sample, with a median of 10 million total readcounts per sample across the transcriptome, and 39,021 transcripts were detected in at least 5% of samples. Comparing PTF to LNM, 6203 transcripts were differentially expressed (p-adj < 0.01). PTF were enriched relative to LNM in gene sets associated with Wnt signaling, hormone signaling, Hippo signaling, KRAS signaling, and the epithelial to mesenchymal transition. Comparing PTF from metastatic patients to non-metastatic patients, 1265 transcripts were differentially expressed (p-adj < 0.01). PTF from metastatic patients were enriched in gene sets associated with cell cycle progression, oxidative phosphorylation, ER stress, fatty acid metabolism, and DNA repair. LNM gene sets were enriched in endoplasmic reticulum (ER) stress and oxidative phosphorylation. The top 500 upregulated genes in malignant tissues were significantly enriched in genes related to androgen and estrogen signaling as expected. We also identified a set of 193 genes whose expression was significantly increased in primary tumor over benign LNs and in LNM over primary tumors. This gene set was significantly enriched in genes related to oxidative phosphorylation and included oncogenes such as PIK3CB, NCOA2, and SCHLAP1. Frequently mutated genes included CSMD3, FAT4, DNAH7, and KMT2C. DNA mutations associated with metastases included PCNX2. Oncogenic pathways associated with metastases in WES analysis included RAS, Notch, Wnt, Hippo, and PI3K pathways. Conclusions: Signaling pathways associated with ER stress, oxidative phosphorylation, metabolism, and cell cycle progression are prominent in LNM of aggressive PCa. PIK3CB, NCOA2, and SCHLAP1 expression are significantly increased in LNM. LNM had higher mutational burdens and were associated with hypermutability pathways, as well as mutations in RAS, Notch, Wnt, Hippo, and PI3K pathway genes. Citation Format: Carlos S. Moreno, Cynthia L. Winham, Emma R. Klein, Yijian Huang, David M. Schuster, Martin G. Sanda, Adeboye O. Osunkoya. Integrated genomic analysis of primary prostate tumor foci and corresponding lymph node metastases identifies pathways associated with metastatic disease [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr A046.

Transcription of WNT Genes in Hematopoietic Niche's Mesenchymal Stem Cells in Multiple Myeloma Patients with Different Responses to Treatment.

Mesenchymal stromal cells (MSCs) are involved in bone tissue remodeling due to their ability to differentiate into osteoblasts and to influence osteoclasts' activity. Multiple myeloma (MM) is associated with bone resorption. During disease progression, MSCs acquire a tumor-associated phenotype, losing their osteogenic potential. The process is associated with impaired osteoblasts/osteoclasts balance. The WNT signaling pathway plays a major role in maintaining the balance. In MM, it functions in an aberrant way. It is not known yet whether the WNT pathway is restored in patients' bone narrow after treatment. The aim of the study was to compare the level of WNT family gene transcription in the bone marrow MSCs of healthy donors and MM patients before and after therapy. The study included healthy donors (n = 3), primary patients (n = 3) and patients with different response status to therapy (bortezomib-containing induction regimens) (n = 12). The transcription of the WNT and CTNNB1 (encoding β-catenin) genes was accessed using qPCR. The mRNA quantity of ten WNT genes, as well as CTNNB1 mRNA encoding β-catenin, a key mediator in canonical signaling, was evaluated. The observed differences between the groups of patients indicated that aberrant functioning of the WNT pathway was retained after treatment. The differences that we detected for WNT2B, WNT9B and CTNNB1 suggested their possible application as prognostic molecular markers.

Antitumor activity of galaxamide involved in cell apoptosis and stemness by inhibiting Wnt/β-catenin pathway in cervical cancer.

Our previous work reported that galaxamide, a cyclopeptide extracted from the seaweed Galaxaura filamentosa, showed antiproliferative activity against HeLa cells by MTT assay. In this study, the growth-inhibitory effects of galaxamide in HeLa cells and xenograft mouse models were investigated. It was found galaxamide significantly inhibited cell growth, colony formation, migration, and invasion and induced cell apoptosis by inhibiting the Wnt signaling pathway in HeLa cells. RNA sequencing revealed that galaxamide regulated stemness by Wnt6 signaling pathway in HeLa cells. By analyzing The Cancer Genome Atlas database, Wnt6 was found to be negatively/positively correlated with stemness- and apoptosis-related genes in human cervical cancer. Cancer stem-like cells (CSCs) isolated and enriched from HeLa cells demonstrated elevated Wnt6 and β-catenin genes compared with nonstem HeLa cells. After galaxamide treatment, CSCs showed abrogation of sphere-forming ability, along with inhibition of stemness-related and Wnt pathway genes. Galaxamide treatment was also accompanied by the induction of apoptosis in HeLa cells, which was consistent with the results in BALB/c nude mice. Our results provide evidence that suppression of stemness by downregulating the Wnt signaling pathway is the molecular mechanism by which galaxamide effectively inhibits cell growth and induces apoptosis in cervical cancer cells.

CDYL reinforces male gonadal sex determination through epigenetically repressing Wnt4 transcription in mice

In mammals, male and female gonads initially develop from bipotential progenitor cells, which can differentiate into either testicular or ovarian cells. The decision to adopt a testicular or ovarian fate relies on robust genetic forces, i.e., activation of the testis-determining gene Sry, as well as a delicate balance of expression levels for pro-testis and pro-ovary factors. Recently, epigenetic regulation has been found to be a key element in activation of Sry. Nevertheless, the mechanism by which epigenetic regulation controls the expression balance of pro-testis and pro-ovary factors remains unclear. Chromodomain Y-like protein (CDYL) is a reader protein for repressive histone H3 methylation marks. We found that a subpopulation of Cdyl-deficient mice exhibited XY sex reversal. Gene expression analysis revealed that the testis-promoting gene Sox9 was downregulated in XY Cdyl-deficient gonads during the sex determination period without affecting Sry expression. Instead, we found that the ovary-promoting gene Wnt4 was derepressed in XY Cdyl-deficient gonads prior to and during the sex-determination period. Wnt4 heterozygous deficiency restored SOX9 expression in Cdyl-deficient XY gonads, indicating that derepressed Wnt4 is a cause of the repression of Sox9. We found that CDYL directly bound to the Wnt4 promoter and maintained its H3K27me3 levels during the sex-determination period. These findings indicate that CDYL reinforces male gonadal sex determination by repressing the ovary-promoting pathway in mice.

Abstract #1400160: Early Onset Osteoporosis Associated with Novel Mutation of WNT1 Gene

Canonical WNT signaling plays a significant role in bone development and maintenance of bone mass. WNT1 gene mutations have been associated with osteogenesis imperfecta type XV in homozygotes and early onset osteoporosis in heterozygotes. We present a case of early onset osteoporosis associated with a novel mutation of the WNT1 gene. A 56-year-old female presented for evaluation of early onset osteoporosis. History was significant for hypothyroidism as well as chronic inflammatory demyelinating polyneuropathy on IVIG infusions. She was diagnosed with low bone density in the setting of a vertebral compression fracture from a diving accident when she was 24. She had mild scoliosis, brittle teeth and 10 fractures as a child and young adult including rib fractures, pelvic fracture and compression fracture of the spine, some of which occurred in the setting of low impact trauma. Her family history was notable for possible osteoporosis in her grandmother. Initial evaluation demonstrated PTH 75 (18 - 80 PG/ML), Calcium 9.5 (8.5 - 10.5 MG/DL ), Albumin 4.1 (3.5 - 5.0 G/DL ), 25-hydroxy vitamin D 44.6 (30.0-100.0 ng/mL), Phosphorus 3.6 (2.4 - 4.8 MG/DL), creatinine 0.59 (0.44 - 1.03 MG/DL), TSH 1.24 (0.350 - 5.500 uIU/ML ), 24-hour urine calcium 133 (50 - 300 MG/24H), late night salivary cortisol 0.066 (less than 0.112 ug/dL), tryptase 6.1 (< 11.0 mcg/L), tissue transglutaminase IgA antibody 2.5 (0-20 units), hemoglobin A1c 4.1 (4.3 - 5.6 % ), IGF-1 112 (50 - 317 ng/mL), Alkaline Phosphorus 62 (34 - 104 IU/L). Dual-energy X-ray absorptiometry (DEXA) scan at the age of 50 showed bone mineral density (BMD) 0.828 g/cm2 with T score of -2.0 at lumbar spine, BMD of 0.593 g/cm2 with T score of -2.3 at left femoral neck and BMD of 0.707 g/cm2 with T score of -1.9. She sustained a low trauma spine fracture at L1 at the age of 55. Subsequent DEXA scan at age 55 showed significant decline of BMD with T score of -3.3 at lumbar spine and T score of -3.0 at the left femoral neck. Due to suspicion for osteogenesis imperfecta, a genetic panel was sent which showed a heterozygous variant in WNT1 gene, c.581T >G (p.L194R). She was treated with oral alendronate 70 mg once weekly for less than one year which was stopped due to gastrointestinal side effects. Follow-up is planned for discussion of additional therapeutic options. Heterozygous mutations in WNT1 have been described in the literature as a cause of early onset osteoporosis. The currently described variant mutation in the WNT1 gene has not been previously reported in literature. In silico analysis using SIFT and PolyPhen2 predict this variant to be deleterious/possibly damaging. Genetic testing and evaluation of bone density in other family members may help further support the clinical importance of this mutation.

Characterization of Wnt genes in Argopecten scallops and their involvement in response to different temperature stresses in “Bohai Red” scallops

Characterization of Wnt genes in Argopecten scallops and their involvement in response to different temperature stresses in “Bohai Red” scallops

Identification of Potential Therapeutic Targets for Plasmablastic Lymphoma Through Gene Expression Analysis: Insights into RAS and Wnt Signaling Pathways

Plasmablastic lymphoma (PBL) is a rare and aggressive B-cell lymphoma with overlapping characteristics with diffuse large B-cell lymphoma (DLBCL) and multiple myeloma. Hyperactive Wnt signaling derails homeostasis and promotes oncogenesis and chemoresistance in DLBCL and multiple myeloma. Evidence suggests active cross-talk between the Wnt and RAS pathways impacting metastasis in solid cancers in which combined targeted therapies show effective results. Recent genomic studies in PBL demonstrated a high frequency of mutations linked with the RAS signaling pathway. However, the role of RAS and Wnt signaling pathway molecule expression in PBL remained unknown. We examined the expression of Wnt and RAS pathway-related genes in a well-curated cohort of PBL. Because activated B cells are considered immediate precursors of plasmablasts in B cell development, we compared this data with activated B-cell type DLBCL (ABC-DLBCL) patients, employing NanoString transcriptome analysis (770 genes). Hierarchical clustering revealed distinctive differential gene expression between PBL and ABC-DLBCL. Gene set enrichment analysis labeled the RAS signaling pathway as the most enriched (37 genes) in PBL, including upregulating critical genes, such as NRAS, RAF1, SHC1, and SOS1. Wnt pathway genes were also enriched (n = 22) by gene set enrichment analysis. Molecules linked with Wnt signaling activation, such as ligands or targets (FZD3, FZD7, c-MYC, WNT5A, WNT5B, and WNT10B), were elevated in PBL. Our data also showed that, unlike ABC-DLBCL, the deranged Wnt signaling activity in PBL was not linked with hyperactive nuclear factor κB and B-cell receptor signaling. In divergence, Wnt signaling inhibitors (CXXC4, SFRP2, and DKK1) also showed overexpression in PBL. The high expression of RAS signaling molecules reported may indicate linkage with gain–in-function RAS mutations. In addition, high expression of Wnt and RAS signaling molecules may pave pathways to explore benefiting from combined targeted therapies, as reported in solid cancer, to improve prognosis in PBL patients.

WNT9A Affects Late-Onset Acute Respiratory Distress Syndrome and 28-Day Survival: Evidence from a Three-Step Multiomics Study.

Late-onset (more than 48 h after ICU admission) acute respiratory distress syndrome (ARDS) is associated with shorter survival time and higher mortality; however, the underlying molecular targets remain unclear. As the WNT gene family is known to drive inflammation, immunity, and tissue fibrosis, all of which are closely related to the pathogenesis and prognosis of ARDS, we aim to investigate the associations of the WNT family with late-onset ARDS and 28-day survival. Genetic (n = 380), epigenetic (n = 185), transcriptional (n = 160), and protein (n = 300) data of patients with ARDS were extracted from the MEARDS (Molecular Epidemiology of ARDS) cohort. We used sure independence screening to identify late onset-related genetic biomarkers and constructed a genetic score on the basis of eight SNPs, which was associated with risk for late-onset ARDS (odds ratio [OR], 2.72; P = 3.81 × 10-14) and survival (hazard ratio [HR], 1.28; P = 0.008). The associations were further externally validated in the iSPAAR (Identification of SNPs Predisposing to Altered Acute Lung Injury Risk) (ORlate onset, 2.49 [P = 0.006]; HRsurvival, 1.87 [P = 0.045]) and MESSI (Molecular Epidemiology of Severe Sepsis in the ICU) (ORlate onset, 4.12 [P = 0.026]; HRsurvival, 1.45 [P = 0.036]) cohorts. Furthermore, we functionally interrogated the six mapped genes of eight SNPs in the multiomics data and noted associations of WNT9A (WNT family member 9A) in epigenetic (ORlate onset, 2.95 [P = 9.91 × 10-4]; HRsurvival, 1.53 [P = 0.011]) and protein (ORlate onset, 1.42 [P = 0.035]; HRsurvival, 1.38 [P = 0.011]) data. The mediation analysis indicated that the effects of WNT9A on ARDS survival were mediated by late onset (HRindirect, 1.12 [P = 0.014] for genetic data; HRindirect, 1.05 [P = 0.030] for protein data). The essential roles of WNT9A in immunity and fibrosis may explain the different trajectories of recovery and dysfunction between early- and late-onset ARDS, providing clues for ARDS treatment.

Division of developmental phases of freshwater leech Whitmania pigra and key genes related to neurogenesis revealed by whole genome and transcriptome analysis

The freshwater leech Whitmania pigra (W. pigra) Whitman (Annelida phylum) is a model organism for neurodevelopmental studies. However, molecular biology research on its embryonic development is still scarce. Here, we described a series of developmental stages of the W. pigra embryos and defined five broad stages of embryogenesis: cleavage stages, blastocyst stage, gastrula stage, organogenesis and refinement, juvenile. We obtained a total of 239.64 Gb transcriptome data of eight representative developmental phases of embryos (from blastocyst stage to maturity), which was then assembled into 21,482 unigenes according to our reference genome sequenced by single-molecule real-time (SMRT) long-read sequencing. We found 3114 genes differentially expressed during the eight phases with phase-specific expression pattern. Using a comprehensive transcriptome dataset, we demonstrated that 57, 49 and 77 DEGs were respectively related to morphogenesis, signal pathways and neurogenesis. 49 DEGs related to signal pathways included 30 wnt genes, 14 notch genes, and 5 hedgehog genes. In particular, we found a cluster consisting of 7 genes related to signal pathways as well as synapses, which were essential for regulating embryonic development. Eight genes cooperatively participated in regulating neurogenesis. Our results reveal the whole picture of W. pigra development mechanism from the perspective of transcriptome and provide new clues for organogenesis and neurodevelopmental studies of Annelida species.

Diversity of Pholcus Spiders (Araneae: Pholcidae) in China’s Lüliang Mountains: An Integrated Morphological and Molecular Approach

Spiders of the genus Pholcus were collected for the first time during an expedition to the Lüliang Mountains in Shanxi Province, North China. Phylogenetic analyses of DNA sequence data from COI, H3, wnt, and 28S genes allowed us to group them into nine well-supported clades. We used morphology and four methods of molecular species delimitation, namely Automatic Barcode Gap Discovery (ABGD), the Generalized Mixed Yule Coalescent (GMYC), Bayesian Poisson Tree Processes (bPTP), and Bayesian Phylogenetics and Phylogeography (BPP), to investigate species boundaries. These integrative taxonomic analyses identified the nine clades as nine distinct species, comprising Pholcus luya Peng & Zhang, 2013 and eight other species new to science: Pholcus jiaocheng sp. nov., Pholcus linfen sp. nov., Pholcus lishi sp. nov., Pholcus luliang sp. nov., Pholcus wenshui sp. nov., Pholcus xiangfen sp. nov., Pholcus xuanzhong sp. nov., and Pholcus zhongyang sp. nov. The species occur in geographic proximity and show many morphological similarities. All of them belong to the P. phungiformes species group. The records from the Lüliang Mountains represent the westernmost distribution limit of this species group.

Abstract 1666: Novel peptides target the β-catenin/TBL1 complex to competitively inhibit dysregulated Wnt signaling in cancer

Abstract Purpose: The canonical Wnt pathway is a stem cell pathway essential during embryogenesis, and functions to direct stem cell fate, renewal, and proliferation. Mutations in this pathway induce overexpression of Wnt genes resulting in a variety of human cancers including breast, colorectal, and ovarian. Canonical Wnt signaling is β-catenin-dependent and when active, β-catenin forms a complex with transducin β-like protein 1 (TBL1) and transducin β-like related protein 1 (TBLR1) to promote gene TCF/LEF-dependent expression downstream the Wnt pathway. Previously we developed a small-molecule inhibitor that disrupts this protein complex to inhibit Wnt signaling and promote a cytotoxic effect in tumor cells. While this approach is effective, our current efforts are focused to develop a peptide-based therapy that targets the β-catenin/TBL1 complex which will result in increased target specificity and less overall toxicity compared to small-molecule counterparts. We have generated a library of peptides corresponding to the binding interface of β-catenin and TBL1 with the aim of developing a specific drug that disrupts the complex. Methods: Selected peptides were tested for their ability to affect the aberrant Wnt pathway on the Wnt-activated colon cancer cell line SW480. Displacement of β-catenin from TBL1 was assessed using a heterologous competition ELISA screen in cell lysate. Hit peptides were identified then tested for specific binding to recombinant TBL1 using an additional ELISA. Further validation to determine associated levels of β-catenin with and without peptide treatment was performed with a co-immunoprecipitation. In situ detection of β-catenin/TBL1 complex dissociation in response to peptide treatment was investigated with a proximity ligation assay (PLA). A ubiquitin enrichment kit was used to measure induced β-catenin degradation. Hit peptides were tested with a viability assay for efficacy in their ability to promote a cytotoxic effect on tumor cells. Transcriptional TCF/LEF activation, which is promoted by β-catenin, was evaluated with TOPFlash in the Wnt activated TCF/LEF reporter HEK293 cell line. Results: We identified a series of peptides that selectively bind TBL1 and displace β-catenin from the complex in Wnt-driven colon cancer cell line SW480 in a dose-dependent manner. Further, we found that treatment with 5 μM peptides in TCF/LEF reporter cells significantly blocks the activity of TCF/LEF, inhibiting downstream transcription of Wnt genes. The disruption of the β-catenin/TBL1 complex results in the ubiquitination of β-catenin. Our findings confirmed this data showing that peptide treatment depletes ubiquitinated β-catenin over time. When these peptides are used to treat Wnt-driven tumor cells, they display strong on-target cytotoxic effects indicating a potential clinical application for the use of these specific peptides as a novel therapeutic. Citation Format: Shelby Rheinschmidt, Trason Thode, Samuel Sampson, Alexis Weston, Tithi Ghosh Halder, Serina Ng, Sydney Adamson, Kate Gutowsky, Mohan Kaadige, Raffaella Soldi, Sunil Sharma. Novel peptides target the β-catenin/TBL1 complex to competitively inhibit dysregulated Wnt signaling in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1666.

Transcriptomic data meta-analysis reveals common and injury model specific gene expression changes in the regenerating zebrafish heart

Zebrafish have the capacity to fully regenerate the heart after an injury, which lies in sharp contrast to the irreversible loss of cardiomyocytes after a myocardial infarction in humans. Transcriptomics analysis has contributed to dissect underlying signaling pathways and gene regulatory networks in the zebrafish heart regeneration process. This process has been studied in response to different types of injuries namely: ventricular resection, ventricular cryoinjury, and genetic ablation of cardiomyocytes. However, there exists no database to compare injury specific and core cardiac regeneration responses. Here, we present a meta-analysis of transcriptomic data of regenerating zebrafish hearts in response to these three injury models at 7 days post injury (7dpi). We reanalyzed 36 samples and analyzed the differentially expressed genes (DEG) followed by downstream Gene Ontology Biological Processes (GO:BP) analysis. We found that the three injury models share a common core of DEG encompassing genes involved in cell proliferation, the Wnt signaling pathway and genes that are enriched in fibroblasts. We also found injury-specific gene signatures for resection and genetic ablation, and to a lower extent the cryoinjury model. Finally, we present our data in a user-friendly web interface that displays gene expression signatures across different injury types and highlights the importance to consider injury-specific gene regulatory networks when interpreting the results related to cardiac regeneration in the zebrafish. The analysis is freely available at: https://mybinder.org/v2/gh/MercaderLabAnatomy/PUB_Botos_et_al_2022_shinyapp_binder/HEAD?urlpath=shiny/bus-dashboard/.

The role of &lt;i&gt;WNT&lt;/i&gt; and &lt;i&gt;HOXA&lt;/i&gt; signaling cascades in the pathogenesis of adenomyosis

BACKGROUND: Adenomyosis is a common gynecological disease with unknown pathogenesis. The HOXA10, HOXA11 and WNT4 genes may play an important role in the pathogenesis of adenomyosis both at the stage of embryonic development and in the postnatal period. The study of their expression in the endometrium of patients with adenomyosis can expand the understanding of the pathogenesis of this disease.&#x0D; AIM: The aim of this work was to study the peculiarity of the WNT4, HOXA10 and HOXA11 gene expression in the eutopic endometrium of patients with isolated adenomyosis.&#x0D; MATERIALS AND METHODS: The study included 38 women: the main group involved patients with isolated adenomyosis established by ultrasound / magnetic resonance imaging (n = 20) and the control group consisted of healthy patients (n = 18). Endometrial sampling was obtained during surgery or by aspiration biopsy at 512 day of the menstrual cycle (proliferative phase) or 2024 day of the menstrual cycle (secretory phase). The expression of the WNT4, HOXA10 and HOXA11 genes in endometrial samples was assessed by a real-time reverse transcription polymerase chain reaction.&#x0D; RESULTS: In the proliferative phase endometrial samples of patients with adenomyosis, a significant increase in the WNT4 (of almost two times), HOXA10 and HOXA11 (of one and a half to two times) gene expression levels was shown compared to the control group. In 88% of patients with adenomyosis, there is a significant increase (up to the level of fourth quartile) in the expression of at least one of these genes, such changes being not typical for the endometrium of women in the control group. In the secretory phase endometrial samples, the expression of the studied genes did not differ from the level characteristic of the corresponding groups in the proliferative phase of the cycle.&#x0D; CONCLUSIONS: The aberrant expression of the WNT4, HOXA10 and HOXA11 genes in the endometrium of patients with adenomyosis indicates a significant role of these genes in the development of the disease and infertility associated with adenomyosis.

2,4-Dipropylphloroglucinol inhibits the growth of human lung and colorectal cancer cells through induction of apoptosis

Scientists are finding the most effective chemotherapeutic agents for the treatment of cancer. In the present study, we evaluated the anticancer mechanism of DPPG, a derivative of DAPG (2,4-diacetylphloroglucinol), for the first time. DPPG and DAPG inhibited 83 and 59% of human colorectal cancer HCT116 cell growth at 40.0µg/ml, and 74 and 57% of human lung cancer A549 cell growth at 10.0µg/ml concentrations respectively. Furthermore, DPPG and DAPG inhibited 97 and 73% colony formation of the HCT116 cells at 20.0µg/ml concentration. DPPG and DAPG induced apoptosis in the HCT116 and A549 cells that was confirmed by Hoechst 33342 and FITC-annexin V staining. This result also revealed that ROS generated in both the HCT116 and A549 cells after treatment with DPPG. However, no ROS production was observed in HCT116 and A549 cells after treatment with DAPG. Both DAPG and DPPG significantly increased the CASP3 protein expression that was detected by staining the cells with the super-view 488-CASP3 substrate. Expression of WNT1 gene was eliminated in DPPG and DAPG treated HCT116. Expression of MAPK1 gene was entirely abolished in DPPG treated cells, whereas a significant decrease was observed for DAPG. An intense band of CASP8 gene product was observed agarose gel for DPPG treated HCT116 cells than DAPG. Molecular docking simulation showed the high binding affinities (≥ 6.5kcal/mol) of DPPG and DAPG with target proteins WNT1, MAPK1, CASP8, and CASP3 in HCT116 cells. This manuscript demonstrated that DAPG and DPPG inhibited lung and colorectal cancer cells by inducing apoptosis. DAPG and DPPG inhibited A549 and HCT116 cells growth by inducing apoptosis.

Genetic variants of IL-10, TGFβ, FGF1, ESR1, MMP1 and WNT4 genes and their association with endometriosis in Iranian women

BackgroundThe complex interaction of genetics, environmental, hormonal, and immunologic factors are involved in the pathogenesis of endometriosis (EMT). This study aimed to investigate the association of controversial and candidate single nucleotide polymorphisms (SNPs) in IL-10 (rs1800872), TGFβ (rs1800469), FGF1 (rs34011), ESR1 (rs9340799; rs2234693), MMP1 (rs1799750), and WNT4 (rs16826658) with EMT in Iranian women with endometriosis. Materials and methodThe seven SNPs in the candidate genes were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique in 101 individuals with EMT and 142 healthy ones as a control group. ResultsOur study revealed a significant association of the FGF1–1385 A/G genotype with the developing EMT in our population (p value = 0.041). However, no significant result was found for other candidate genotypes and alleles in susceptibility to EMT. ConclusionFGF1–1385 A/G SNP genotypes might be correlated with the pathogenesis of EMT in Iranian women.