Development Of Targeted Therapies Research Articles

SNP Analysis of the SOD1 gene involved in Amyotrophic Lateral Sclerosis (ALS)

ALS is a rare neurodegenerative disease causing the loss of motor neurons in the brain and spinal cord. Mutations in over forty genes, particularly the Superoxide Dismutase 1 (SOD1) gene, are linked to ALS. This study analyses the missense or non-synonymous SNPs (nsSNPs) in the coding region of SOD1 using computational tools to predict the variations in structure and function of protein. The variants with most pathogenicity and clinical significance were further analyzed for stability and found to destabilize the protein structure based on Gibbs Free Energy (ddG) values. These 10 variants were modelled and validated, found occurring mostly in the conserved regions. Among these, 9 nsSNPs reported previously, while 1 novel nsSNP (F46S) was identified, characterized by reduced size and hydrobhobicity in comparison to the wild-type residue. This comprehensive study improves our understanding of the effects of SOD1 SNPs in ALS, thereby aiding in targeted therapies development.

Development of FAP-targeted theranostics discovered by next-generation sequencing-augmented mining of a novel immunized VNAR library.

Cancer-associated fibroblasts (CAFs) in the stroma of solid tumors promote an immunosuppressive tumor microenvironment (TME) that drives resistance to therapies. The expression of the protease fibroblast activation protein (FAP) on the surface of CAFs has made FAP a target for development of therapies to dampen immunosuppression. Relatively few biologics have been developed for FAP and none have been developed that exploit the unique engagement properties of Variable New Antigen Receptors (VNARs) from shark antibodies. As the smallest binding domain in nature, VNARs cleverage unique geometries and recognize epitopes conventional antibodies cannot. By directly immunizing a nurse shark with FAP, we created a large anti-FAP VNAR phage display library. This library allowed us to identify a suite of anti-FAP VNARs through traditional biopanning and also by an in silico approach that did not require any prior affinity-based enrichment in vitro. We investigated four VNAR-Fc fusion proteins for theranostic properties and found that all four recognized FAP with high affinity and were rapidly internalized by FAP-positive cells. As a result, the VNAR-Fc constructs were effective antibody-drug conjugates in vitro and were able to localize to FAP-positive xenografts in vivo. Our findings establish VNAR-Fc constructs as a versatile platform for theranostic development that could yield innovative cancer therapies targeting the TME.

NEK2 promotes colorectal cancer progression by activating the TGF-β/Smad2 signaling pathway

Colorectal cancer (CRC) is a prevalent malignancy with poor patient survival, and NIMA-associated kinase 2 (NEK2) has been implicated in the pathogenesis and progression of various cancers, including CRC. This study aimed to investigate the impact of NEK2 on CRC cell functionality and its interaction with the TGF-β/Smad signaling pathway. NEK2 expression in CRC tissues and cell lines was assessed, and its association with patient survival was analyzed. Functional assays, including NEK2 knockdown via lentiviral infection, RT-qPCR, Western blotting, CCK-8 assay, Transwell migration, invasion assays, and goblet cell formation assays, were employed to evaluate NEK2′s effects on CRC cell proliferation, migration, invasion, and stemness. Mechanistic studies explored the TGF-β/Smad2 signaling pathway, utilizing co-immunoprecipitation (Co-IP) and protein interaction analyses. In vivo experiments further evaluated NEK2′s role in tumor initiation, metastasis, and chemoresistance. NEK2 was found to be upregulated in CRC tissues and correlated with poor survival. NEK2 knockdown inhibited CRC cell behaviors, while NEK2 activated the TGF-β/Smad2 signaling pathway through Smad2/3 phosphorylation. Overexpression of Smad2/3 reversed NEK2 knockdown effects, confirming the importance of this pathway in CRC. In vivo, NEK2 promoted tumor initiation, metastasis, and chemoresistance, effects partially reversed by Smad2/3 overexpression. These findings reveal the critical role of NEK2 in CRC progression and underscore its potential as a therapeutic target, offering new insights into the molecular mechanisms driving CRC and informing targeted therapy development.

LncRNA-Associated ceRNA Network Revealing the Potential Regulatory Roles of Ferroptosis and Immune Infiltration in Osteosarcoma as well as Construction of the Prognostic Model.

Osteosarcoma (OS) is the most common primary bone malignancy in the world. Increasing studies indicate that long non-coding RNAs (lncRNAs) are involved in ferroptosis and OS progression. Therefore, this study aims to identify ferroptosis- related lncRNAs (frlncRNAs), explore potential competing endogenous RNA (ceRNA) networks, and establish a new model for predicting OS prognosis. Firstly, we downloaded data from The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), University of California, Santa Cruz (UCSC), and FerrDB, and screened for differentially expressed FRlncRNAs (DEFRlncRNAs) between OS patients and healthy controls. Then, we constructed the ceRNA network using the Lncbase 3.0, starBase, miRDB, miRTarBase, and TargetScan databases. Subsequently, prognosis- related DEFRlncRNAs were selected through Cox analysis, and a prognostic model was constructed. Next, the proportions of different immune cells in high and low-risk groups were quantified and evaluated using the "CIBERSORT" algorithm. Finally, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on prognosis-related DEFRlncRNAs to identify topranked biological processes and pathways. We identified 247 DEFRlncRNAs and constructed the ceRNA network comprising 37 lncRNAs, 84 microRNAs (miRNAs), and 865 messenger RNAs (mRNAs). Subsequently, we obtained 8 prognosis-related DEFRlncRNAs (AL645728.1, AL161785.1, LINC00539, AL590764.1, OLMALINC, AC110995.1, AC091180.2, and AL160006.1) and constructed a prognostic model, where metastasis and risk score were identified as important clinical factors for predicting OS prognosis. Additionally, only OLMALINC and AL160006.1 had corresponding target miRNAs in the prognosis-related ceRNA network. Lastly, we revealed the infiltration proportions of different immune cells in OS, with higher proportions of macrophages (M0 and M2 subgroups) and T cells (T cells CD4 memory resting and T cells CD8) observed. This study explored the ferroptosis-related lncRNA-miRNA-mRNA regulatory network in OS, constructed a ferroptosis-related prognostic model, and characterized its association with immune infiltration, providing new insights into the pathological mechanisms and targeted therapy development for OS.

Application of Single-Cell Genomics to Animal Models ofPeriodontitis and Peri-Implantitis.

This narrative review aims to synthesize current knowledge on integrating single-cell genomics technologies with animal models of periodontitis and peri-implantitis. Single-cell RNA sequencing (scRNAseq) reveals cellular heterogeneity and specific cell roles in periodontitis and peri-implantitis, overcoming the limitations of bulk RNA sequencing. Under controlled conditions and genetic manipulation, animal models facilitate studying disease progression, gene functions and systemic disease links, aiding targeted therapy development. Knockout models have started to elucidate the impact of genetic mutations on periodontal disease and host responses. scRNAseq in animal models has been used to examine connections between periodontitis and systemic diseases, revealing altered immune environments and cellular interactions. Emerging studies are now applying these methods to animal models of peri-implantitis. Integrating these datasets into single-cell and spatially resolved atlases will enable future meta-analyses, providing deeper insights into disease mechanisms considering factors such as sex, strain, and age. Integrating scRNAseq with animal models advances the understanding of periodontitis and peri-implantitis pathogenesis and precision therapies. The combined use of single-cell and spatial genomics and scRNAseq will further enhance data insights significantly for drug discovery and preclinical testing, making these technologies pivotal in validating animal models and translating findings into clinical practice.

HSP and CD279 gene expression as candidate biomarkers in symptomatic LGLL patients

The clinical presentation of T-cell large granular lymphocytic leukemia (T-LGLL) is extremely variable: 30% of patients have neutropenia with no associated symptoms, others present with bacterial infections and sepsis may occur. Tools to predict patient outcome are lacking. Stemming from preliminary results obtained by single cell-RNAseq we investigated by qPCR HSP and IFIT gene families in 27 LGLL patients (23T-LGLL and 4 NK-LGLL), including 11 with neutropenia and/or thrombocytopenia and 16 asymptomatic for the disease. HSP90AA1 and HSPA1B, among HSP family and CD279 exhibited a significantly higher expression in CD3 + CD57 + sorted cells of symptomatic LGLL patients compared to asymptomatic patients and healthy controls. Also, monocytes derived from symptomatic LGLL patients expressed high levels of CCL3, CCL4 and CCL5 mRNA and of IL-1β, IL-6, TNF, and PD-L1 mRNA, thus confirming a pro-inflammatory cytokine profile reminiscent of a non-classical phenotype. Overall, these data provide a rationale for considering HSP and CD279 genes as potential biomarkers for distinguishing symptomatic LGLL patients from asymptomatic ones, emphasizing the importance of further research to explore their implications for targeted therapy development.

Calcific aortic stenosis: omics-based target discovery and therapy development.

Calcific aortic valve disease (CAVD) resulting in aortic stenosis (AS) is the most common form of valvular heart disease, affecting 2% of those over age 65. Those who develop symptomatic severe AS have an average further lifespan of <2 years without valve replacement, and three-quarters of these patients will develop heart failure, undergo valve replacement, or die within 5 years. There are no approved pharmaceutical therapies for AS, due primarily to a limited understanding of the molecular mechanisms that direct CAVD progression in the complex haemodynamic environment. Here, advances in efforts to understand the pathogenesis of CAVD and to identify putative drug targets derived from recent multi-omics studies [including (epi)genomics, transcriptomics, proteomics, and metabolomics] of blood and valvular tissues are reviewed. The recent explosion of single-cell omics-based studies in CAVD and the pathobiological and potential drug discovery insights gained from the application of omics to this disease area are a primary focus. Lastly, the translation of knowledge gained in valvular pathobiology into clinical therapies is addressed, with a particular emphasis on treatment regimens that consider sex-specific, renal, and lipid-mediated contributors to CAVD, and ongoing Phase I/II/III trials aimed at the prevention/treatment of AS are described.

Advancements in Dental Bioreactor Design: A Comprehensive Approach for Application in Dentistry.

The protocol introduces a novel multi-chamber bioreactor tailored for ex-vivo cell culture in dentistry research, emulating the 3D dental environment to propel research in dental applications. Constructed primarily from a polymeric material with a sophisticated 3D design, the bioreactor securely holds teeth structures within sealed chambers, enabling controlled perfusion of culture medium crucial for cell growth through a singular entry and exit point. An integrated electronic system manages flow and pressure, ensuring precise control over environmental conditions. This technology facilitates cell cultivation under conditions closely resembling natural tooth microenvironments, offering opportunities for varied studies from understanding cellular behavior in dental contexts to targeted therapy development. The bioreactor's amalgamation of polymeric components, 3D design, and electronic controls enhances adaptability and accuracy, rendering it a valuable asset in dental research. The report comprehensively delineates the bioreactor's design, operations, and potential applications, showcasing its significant contributions to dental research and regenerative medicine. By amalgamating advanced technologies, this bioreactor emerges as a pivotal tool for investigating cellular processes within dental structures, paving the way for scientific exploration and therapeutic advancements.

WONOEP appraisal: Targeted therapy development for early onset epilepsies.

The early onset epilepsies encompass a heterogeneous group of disorders, some of which result in drug-resistant seizures, developmental delay, psychiatric comorbidities, and sudden death. Advancement in the widespread use of targeted gene panels as well as genome and exome sequencing has facilitated the identification of different causative genes in a subset of these patients. The ability to recognize the genetic basis of early onset epilepsies continues to improve, with de novo coding variants accounting for most of the genetic etiologies identified. Although current disease-specific and disease-modifying therapies remain limited, novel precision medicine approaches, such as small molecules, cell therapy, and other forms of genetic therapies for early onset epilepsies, have created excitement among researchers, clinicians, and caregivers. Here, we summarize the main findings of presentations and discussions on novel therapeutic strategies for targeted treatment of early onset epilepsies that occurred during the Workshop on Neurobiology of Epilepsy (WONOEP XVI, Talloires, France, July 2022). The presentations discussed the use of chloride transporter inhibitors for neonatal seizures, targeting orexinergic signaling for childhood absence epilepsy, targeting energy metabolism in Dravet syndrome, and the role of cannabinoid receptor type 2, reversible acetylcholinesterase inhibitors, cell therapies, and RNA-based therapies in early life epilepsies.

Exploring the molecular profile of localized colon cancer: insights from the AIO Colopredict Plus registry.

Understanding the mutational landscape of colon cancer (CC) is crucial for targeted therapy development. Microsatellite instability (MSI-H), rat sarcoma (RAS), and B-Raf proto-oncogene, serine/threonine kinase (BRAF) mutations (MT) are pivotal markers. Further investigation into clinicopathological features of RAS and BRAF MT in microsatellite stable (MSS) and MSI-H tumors is warranted. A retrospective analysis of 4883 localized CC patients (pts.) was conducted. Molecular profiling assessed MSI, KRAS, NRAS, and BRAF MT. Correlation with clinicopathological data employed ANOVA and Chi-square tests. Disease-free survival (DFS) and overall survival (OS) were analyzed adjusting for age, gender, sidedness, UICC stage, Charlson Comorbidity Index (CCI). A Cox model incorporated all variables as covariates. This analysis included 4883 pts. (2302 female/2572 male, 3865 (79.2%) MSS, 1018 (20.8%) MSI-H). MSS pts. had more All-Wild Type (WT), KRAS MT, and NRAS MT tumors vs. MSI-H pts. (42.1% vs. 21.1%; 39.8% vs. 15.4%; 3.6% vs. 0.7%; p<0.001 for each). BRAF MT tumors (95.5% BRAF V600E MT) were more prevalent in MSI-H individuals (62.8% vs. 8.1%, p<0.001). KRAS and BRAF MT tumors were more frequently right-sided, while BRAF MT tumors were associated with female gender, advanced disease stage, lymph node positivity, and poorer differentiation in the MSS subset (p<0.001). Common KRAS mutations included p.G12D (30.44%) and p.G12V (21.3%) in MSS and p.G13D (28.9%) and p.G12D (22.37%) in MSI-H. NRAS MT tumors were dominated by codon 61 mutations (51.7%). Survival analysis revealed worst prognosis in BRAF MT MSS tumors (DFS: HR 1.74 (95% CI 1.15-2.62, p=0.009; OS: HR 1.61 (95% CI 0.99-2.6), p=0.055). The 3-years DFS and 5-years OS rates were lowest in this subset (61.6% and 57.7% respectively). These findings highlight the complex interplay between molecular subtypes, clinicopathological features, and survival outcomes in early CC. Further research is needed to elucidate underlying mechanisms and develop personalized treatment strategies.

Immunoglobulin A nephropathy is not what it used to be!

Immunoglobulin A (IgA) nephropathy is the most common primary glomerulonephritis worldwide and thought to be a benign disease. Recent literature review would suggest otherwise, with outcomes remaining generally poor. Few patients do not progress to kidney failure in their lifetime even in population groups previously thought to be low risk. The previously established treatment approach predominantly focuses on supportive care through renin-angiotensin-aldosterone system inhibition and steroid-based immunosuppression in selected cases. However, recent advances in the understanding of the pathogenic mechanisms underlying this disease process have allowed for the development of new target therapies. This brief review provides an overview of improved understanding of IgA nephropathy and novel therapeutic strategies.

Evaluation of CD96 Expression in Acute Myeloid Leukemia Patients: Relation to Prognosis and Induction Chemotherapy

Abstract Background In acute myeloid leukemia (AML), leukemic stem cells (LSCs) form a distinct group of leukemia cells showing stem cell-like features like self-renewal and proliferation. These potent cells significantly contribute to chemotherapy resistance, a major obstacle in AML therapy. LSCs express the characteristic phenotype CD34+/CD38- and have several specific markers, including CD33, CD96, CD123, CD90 and Death associated protein kinase (DAPK). Successful LSC eradication demands a combination of approaches, including targeting LSC-specific surface molecules. Objective To assess the expression of CD96 in AML patients at initial diagnosis and to correlate it with prognostic factors and response to induction chemotherapy. Materials and Methods This Case control study was conducted on 40 newly diagnosed AML patients in addition to 10 controls at Ain Shams University Hospitals between March 2022 and March 2023. Multicolor flowcytometry was employed to analyze CD96 expression in the CD34+/CD38- cell population of AML patients. The study further investigated the correlation between CD96 expression and both prognostic factors and response to induction chemotherapy. Results CD96 expression was significantly higher in AML patients group than control group (p-value = 0.008). CD96 was positive in 60% of AML patients and there was a highly significant association between CD96 expression and response to induction chemotherapy (p-value = &amp;lt;0.001). However, there was no statistically significant relation between CD96 expression with laboratory findings and standard prognostic factors. Conclusion Our study identified CD96 as a promising marker for LSCs in AML patients. Its frequent expression in the CD34+/CD38- LSC population and association with lower response rate to induction chemotherapy suggest its potential as a valuable tool for LSC identification, targeted therapy development and improved treatment outcome prediction in AML.

Cluster analysis of blood biomarkers to identify molecular patterns in pulmonary fibrosis: assessment of a multicentre, prospective, observational cohort with independent validation

Pulmonary fibrosis results from alveolar injury, leading to extracellular matrix remodelling and impaired lung function. This study aimed to classify patients with pulmonary fibrosis according to blood biomarkers to differentiate distinct disease patterns, known as endotypes. In this cluster analysis, we first classified patients from the PROFILE study, a multicentre, prospective, observational cohort of individuals with incident idiopathic pulmonary fibrosis or non-specific interstitial pneumonia in the UK (Nottingham University Hospitals, Nottingham; and Royal Brompton Hospital, London). 13 blood biomarkers representing extracellular matrix remodelling, epithelial stress, and thrombosis were measured by ELISA in the PROFILE study. We classified patients by unsupervised consensus clustering. To evaluate generalisability, a machine learning classifier trained on biomarker signatures derived from consensus clustering was applied to a replication dataset from the Australian Idiopathic Pulmonary Fibrosis Registry (AIPFR). Biomarker associations with mortality and change in percentage of predicted forced vital capacity (FVC%) were assessed, adjusting for age, gender, baseline FVC%, and antifibrotic treatment and steroid treatment before and after baseline. Mortality risk associated with the clusters in the PROFILE cohort was evaluated with Cox proportional hazards models, and mixed-effects models were used to analyse how clustering was associated with longitudinal FVC% in the PROFILE and AIPFR cohorts. 455 of 580 participants from the PROFILE study (348 [76%] men and 107 [24%] women; mean age 72·4 years [SD 8·3]) were included in the analysis. Within this group, three clusters were identified based on blood biomarkers. A basement membrane collagen (BM) cluster (n=248 [55%]) showed high concentrations of PRO-C4, PRO-C28, C3M, and C6M, whereas an epithelial injury (EI) cluster (n=109 [24%]) showed high concentrations of MMP-7, SP-D, CYFRA211, CA19-9, and CA-125. The third cluster (crosslinked fibrin [XF] cluster; n=98 [22%]) had high concentrations of X-FIB. In the replication dataset (117 of 833 patients from AIPFR; 87 [74%] men and 30 [26%] women; mean age 72·9 years [SD 7·9]), we identified the same three clusters (BM cluster, n=93 [79%]; EI cluster, n=8 [7%]; XF cluster, n=16 [14%]). These clusters showed similarities with clusters in the PROFILE dataset regarding blood biomarkers and phenotypic signatures. In the PROFILE dataset, the EI and XF clusters were associated with increased mortality risk compared with the BM cluster (EI vs BM: adjusted hazard ratio [HR] 1·88 [95% CI 1·42-2·49], p<0·0001; XF vs BM: adjusted HR 1·53 [1·13-2·06], p=0·0058). The EI cluster showed the greatest annual FVC% decline, followed by the BM and XF clusters. A similar FVC% decline pattern was observed in these clusters in the AIPFR replication dataset. Blood biomarker clustering in pulmonary fibrosis identified three distinct blood biomarker signatures associated with lung function and prognosis, suggesting unique pulmonary fibrosis biomarker patterns. These findings support the presence of pulmonary fibrosis endotypes with the potential to guide targeted therapy development. None.

Deletion of cyclin dependent kinase inhibitor 2a gene as a marker of oropharyngeal carcinomas non-associated with human papillomavirus and its prognostic value

Patients with human papilloma virus associated oropharyngeal squamous cell carcinoma generally have better treatment outcomes and prognosis compared to those with non-papillomavirus-associated oropharyngeal squamous cell carcinoma. However, prognostic evaluation for non-papillomavirus-associated oropharyngeal squamous cell carcinoma remains a problem that could be solved through the molecular mechanisms of squamous cell carcinoma for the purpose of further development of target therapies. Detection of cyclin dependent kinase inhibitor 2a gene deletion in oropharyngeal squamous cell carcinomas can have clinical significance as it may serve as a prognostic marker and potentially guide treatment decisions. To investigate and analyze cyclin dependent kinase inhibitor 2a gene alterations in oropharyngeal squamous cell carcinoma comparing with clinical data (age of the patient, TNM stage), their histological features and occurrence of papillomavirus infection markers (p16 expression). Formalin-fixed and paraffin-embedded samples after transoral radical surgery of oropharyngeal tumors from 26 male patients with average age 57.35±10.33 years were studied. Histological, immunohistochemical analyses and fluorescent in-situ hybridization were performed to assess histological features, p16 expression, and cyclin dependent kinase inhibitor 2a gene gene abnor­malities respectively. Homozygous deletion of cyclin dependent kinase inhibitor 2a gene was statistically analyzed and compared with p16 expression, age, and occurrence of nodal metastases in investigated patients. Our study demonstrated that the patients with non-papillomavirus-oropharyngeal squamous cell carcinoma with cyclin dependent kinase inhibitor 2a gene homozygous deletion had the highest risk of the nodal metastases development. Our findings suggest that not only detection of the loss of p16 expression, but also the evaluation of homozygous cyclin dependent kinase inhibitor 2a gene deletion might be predictive of worse outcome specifically in oropharyngeal squamous cell carcinomas.

Exosomes from umbilical cord mesenchymal stromal cells promote the collagen production of fibroblasts from pelvic organ prolapse.

Pelvic organ prolapse (POP) involves pelvic organ herniation into the vagina due to pelvic floor tissue laxity, and vaginal structure is an essential factor. In POP, the vaginal walls exhibit abnormal collagen distribution and decreased fibroblast levels and functions. The intricate etiology of POP and the prohibition of transvaginal meshes in pelvic reconstruction surgery present challenges in targeted therapy development. Human umbilical cord mesenchymal stromal cells (hucMSCs) present limitations, but their exosomes (hucMSC-Exo) are promising therapeutic tools for promoting fibroblast proliferation and extracellular matrix remodeling. To investigate the effects of hucMSC-Exo on the functions of primary vaginal fibroblasts and to elucidate the underlying mechanism involved. Human vaginal wall collagen content was assessed by Masson's trichrome and Sirius blue staining. Gene expression differences in fibroblasts from patients with and without POP were assessed via RNA sequencing (RNA-seq). The effects of hucMSC-Exo on fibroblasts were determined via functional experiments in vitro. RNA-seq data from fibroblasts exposed to hucMSC-Exo and microRNA (miRNA) sequencing data from hucMSC-Exo were jointly analyzed to identify effective molecules. In POP, the vaginal wall exhibited abnormal collagen distribution and reduced fibroblast 1 quality and quantity. Treatment with 4 or 6 μg/mL hucMSC-Exo suppressed inflammation in POP group fibroblasts, stimulated primary fibroblast growth, and elevated collagen I (Col1) production in vitro. High-throughput RNA-seq of fibroblasts treated with hucMSC-Exo and miRNA sequencing of hucMSC-Exo revealed that abundant exosomal miRNAs downregulated matrix metalloproteinase 11 (MMP11) expression. HucMSC-Exo normalized the growth and function of primary fibroblasts from patients with POP by promoting cell growth and Col1 expression in vitro. Abundant miRNAs in hucMSC-Exo targeted and downregulated MMP11 expression. HucMSC-Exo-based therapy may be ideal for safely and effectively treating POP.

Exosomes from umbilical cord mesenchymal stromal cells promote the collagen production of fibroblasts from pelvic organ prolapse

BACKGROUND Pelvic organ prolapse (POP) involves pelvic organ herniation into the vagina due to pelvic floor tissue laxity, and vaginal structure is an essential factor. In POP, the vaginal walls exhibit abnormal collagen distribution and decreased fibroblast levels and functions. The intricate etiology of POP and the prohibition of transvaginal meshes in pelvic reconstruction surgery present challenges in targeted therapy development. Human umbilical cord mesenchymal stromal cells (hucMSCs) present limitations, but their exosomes (hucMSC-Exo) are promising therapeutic tools for promoting fibroblast proliferation and extracellular matrix remodeling. AIM To investigate the effects of hucMSC-Exo on the functions of primary vaginal fibroblasts and to elucidate the underlying mechanism involved. METHODS Human vaginal wall collagen content was assessed by Masson’s trichrome and Sirius blue staining. Gene expression differences in fibroblasts from patients with and without POP were assessed via RNA sequencing (RNA-seq). The effects of hucMSC-Exo on fibroblasts were determined via functional experiments in vitro . RNA-seq data from fibroblasts exposed to hucMSC-Exo and microRNA (miRNA) sequencing data from hucMSC-Exo were jointly analyzed to identify effective molecules. RESULTS In POP, the vaginal wall exhibited abnormal collagen distribution and reduced fibroblast 1 quality and quantity. Treatment with 4 or 6 μg/mL hucMSC-Exo suppressed inflammation in POP group fibroblasts, stimulated primary fibroblast growth, and elevated collagen I (Col1) production in vitro . High-throughput RNA-seq of fibroblasts treated with hucMSC-Exo and miRNA sequencing of hucMSC-Exo revealed that abundant exosomal miRNAs downregulated matrix metalloproteinase 11 (MMP11) expression. CONCLUSION HucMSC-Exo normalized the growth and function of primary fibroblasts from patients with POP by promoting cell growth and Col1 expression in vitro . Abundant miRNAs in hucMSC-Exo targeted and downregulated MMP11 expression. HucMSC-Exo-based therapy may be ideal for safely and effectively treating POP.

The Evolving Scenario of ES-SCLC Management: From Biology to New Cancer Therapeutics.

Small cell lung cancer (SCLC) is an aggressive neuroendocrine carcinoma accounting for 15% of lung cancers with dismal survival outcomes. Minimal changes in therapy and prognosis have occurred in SCLC for the past four decades. Recent progress in the treatment of extensive-stage disease (ES-SCLC) has been marked by incorporating immune checkpoint inhibitors (ICIs) into platinum-based chemotherapy, leading to modest improvements. Moreover, few second-line-and-beyond treatment options are currently available. The main limitation for the molecular study of SCLC has been the scarcity of samples, because only very early diseases are treated with surgery and biopsies are not performed when the disease progresses. Despite all these difficulties, in recent years we have come to understand that SCLC is not a homogeneous disease. At the molecular level, in addition to the universal loss of retinoblastoma (RB) and TP53 genes, a recent large molecular study has identified other mutations that could serve as targets for therapy development or patient selection. In recent years, there has also been the identification of new genetic subtypes which have shown us how intertumor heterogeneity exists. Moreover, SCLC can also develop intratumoral heterogeneity linked mainly to the concept of cellular plasticity, mostly due to the development of resistance to therapies. The aim of this review is to quickly present the current standard of care of ES-SCLC, to focus on the molecular landscapes and subtypes of SCLC, subsequently present the most promising therapeutic strategies under investigation, and finally recap the future directions of ongoing clinical trials for this aggressive disease which still remains a challenge.

Characteristics and outcomes of acute myeloid leukaemia patients with baseline CD7 expression.

Targeted therapy development for acute myeloid leukaemia (AML) requires an understanding of specific expression profiles. We collected flow cytometry data on 901 AML patients and recorded aberrant CD7 expression on leukaemic blasts. 263 (29.2%) had blasts positive for CD7. CD7+ AML was more likely to be adverse risk (64.6% vs. 55.6%, p = 0.0074) and less likely to be favourable risk (15.2% vs. 24.1%, p = 0.0074) by European LeukemiaNet 2022 criteria. Overall survival was inferior (11.9 [95% CI, 9.7-15.9] vs. 19.0 months [95% CI, 16.1-23.0], p = 0.0174). At relapse, 30.4% lost and 19.0% gained CD7, suggesting moderate instability over time.

Molecular Pathology of Hematologic Malignancies

Abstract Hematological malignancies represent a heterogeneous group of diseases that may have overlapping clinical manifestations. Successful and optimal management depends on early and accurate diagnosis of the disease. Differential diagnosis therefore requires methods of morphology, immunohistochemistry, flow cytometry, and also molecular and cytogenetic examinations. Molecular diagnostic techniques are becoming more accurate and sophisticated, which is why nowadays hematopathological diagnosis relies heavily on molecular and cytogenetic analyses. They are beneficial not only for diagnosis, but also for evaluating prognostic and risk markers, as well as treatment monitoring. This article presents an overview of selected hematological malignancies – mature lymphoid neoplasms, multiple myelomas, myeloproliferative neoplasms, myelodysplastic syndromes, and acute myeloid leukemias; their molecular pathology, risk and prognostic markers. Understanding of the biologic basis leds to a targeted therapy development for the treatment of these diseases.

COL8A1 Regulates Esophageal Squamous Carcinoma Proliferation and Invasion Through PI3K/AKT Pathway.

Esophageal squamous carcinoma (ESCC) is a gastrointestinal malignancy with a high mortality, but the tumorigenesis is still unclear, restricting the target therapy development of ESCC. We explored the role of COL8A1 in ESCC development. Tissue microarrays were used to investigate the expression level of COL8A1 in ESCC tissues. The association between COL8A1 and the overall survival of ESCC patients was assessed. The effect of differential COL8A1 expression on tumor growth was investigated by the xenograft model. The regulation of COL8A1 on tumor growth, migration, and invasion was studied by using ESCC cell lines. The signal transduction pathways involved in COL8A1 were bioinformatically profiled and validated. The COL8A1 was significantly expressed in cancerous tissues and was associated with poor prognosis in patients with ESCC. In vivo, the tumor growth obviously declined after inhibition of the COL8A1 expression. The abilities of cell proliferation and invasion were both decreased when the expression of COL8A1 was knockdown in ESCC cell line. Furthermore, we found the inactivation of the PI3K/AKT pathway that was mediated by knockdown of COL8A1 in ESCC cells, which was reversed with COL8A1 overexpression, whereas the cell proliferation and invasion ability were restored. This is the first report that COL8A1 promote ESCC progression, which hopefully will provide a theoretical basis for clinical targeting of ESCC.