Shafaat Ahmed Rabbani

Background Methyl-CpG-binding domain protein 2 (MBD2) is a key epigenetic regulator implicated in tumorigenesis by repressing tumor suppressor genes by recognizing DNA methylation marks and recruiting specific histone-modifying enzymes and chromatin remodeling complexes. Although KCC-07 has been identified as a potent selective MBD2 inhibitor, its cytotoxic effects on various cancer cells remain largely unexplored. In the current study, we have examined the anti-proliferative and cytotoxic activities of KCC-07 on breast cancer (MCF-7), prostate cancer (PC-3), hepatocellular carcinoma (Huh-7), and osteosarcoma (U2-OS) cell lines, along with human skin fibroblasts (HFF-1) as a non-malignant control. Methods and Results Treatment of these cells with KCC-07 induced dose- and time-dependent cytotoxicity, notably reducing viability in Huh-7 and PC-3 cells. Flow cytometry analyses revealed that KCC-07 primarily triggered necrosis in Huh-7 and apoptosis in PC-3 cells. Furthermore, KCC-07 substantially downregulated MBD2-associated oncogenic targets such as WNT1, CCND1, and ERK1/2 in Huh-7 and PC-3 cells without altering MBD2 transcription. Conclusion These findings suggest that KCC-07 elicited a considerable cytotoxicity on cancer cells, likely modulating the expression of MBD2-related genes, highlighting its potential as a candidate for cancer therapy. Graphical Abstract

The urokinase-type plasminogen activator receptor (uPAR) plays a pivotal role in regulating extracellular proteolysis, cell migration, immune responses, and tissue remodeling across diverse physiological and pathological contexts. This review provides detailed insights into the structure of uPAR, ligand interactions, and signaling mechanisms, emphasizing its central function in cancer progression, including tumor invasion, metastasis, angiogenesis, and modulation of the tumor microenvironment. We also summarize the involvement of uPAR as a key player in cardiovascular, infectious, and neurological diseases, where it contributes to inflammation, tissue damage, and disease progression. However, translational gaps remain, most notably inconsistent assay harmonization (especially for suPAR), uncertain context-specific cut-offs and patient-selection criteria and limited multicenter validation for uPAR-targeted imaging and therapeutics. This review addresses these gaps by synthesizing cross-disease evidence to clarify clinical use cases and outline practical selection frameworks. Furthermore, we discuss the clinical potential of uPAR as a diagnostic and prognostic biomarker in diverse disease contexts, along with recent advances in therapeutic strategies targeting uPAR.

Background Melanoma is one of the most serious skin cancers worldwide, often progressing without obvious symptoms. Early detection is crucial to enable timely intervention, reducing morbidity and mortality. However, no standardized high-throughput test exists for non-invasive melanoma detection. Methods This diagnostic study aimed to develop and validate a high-throughput targeted DNA methylation-based test for detecting melanoma in cell-free DNA (cfDNA) from plasma and predicting response to immune checkpoint inhibitors (ICIs). A multiplexed next-generation sequencing assay targeting these five melanoma-specific DNA regions, named epiMelanoma, was developed and tested on a clinical cohort of 199 participants, including 121 melanoma patients (stages I-IV) from oncology clinics and 78 healthy donors sourced from biorepositories. Plasma cfDNA was collected from 187 participants, and biopsies from 35 patients. Results Here we show that epiMelanoma shows high classification accuracy in both biopsies and plasma cfDNA. First, four DNA regions categorically methylated across diverse cancers, including melanoma, but unmethylated in other tissues, were identified using TCGA and GEO datasets (n = 160), showing high classification accuracy (AUC = 0.9987, sensitivity 98.67%, specificity 100%). A fifth melanoma-specific region was discovered (cg04652957, sensitivity 100%, specificity 90.91%) and validated in 5479 samples. In clinical samples, sensitivity for early- and late-stage melanoma was 27% and 60%, respectively, with high specificity (98.15%). Sensitivity in biopsies exceeded 90%. Furthermore, a significant correlation was observed between the methylation signature and response to ICIs (p = 0.012), with lower M-scores associated with improved overall survival (log-rank p = 0.001, Kaplan–Meier). Conclusions The epiMelanoma assay represents a promising non-invasive tool for early melanoma detection and personalized treatment, by enabling timely intervention and identifying patients most likely to benefit from immunotherapy, and therefore, potentially improving patient outcomes and reducing healthcare burdens.

Methyl-CpG-binding domain protein 2 (Mbd2), a reader of DNA methylation, has been implicated in different types of malignancies, including breast cancer. However, the exact role of Mbd2 in various stages of breast cancer growth and progression in vivo has not been determined. To test whether Mbd2 plays a causal role in mammary tumor growth and metastasis, we performed genetic knockout (KO) of Mbd2 in MMTV-PyMT transgenic mice and compared mammary tumor progression kinetics between the wild-type (PyMT-Mbd2^+/+) and KO (PyMT-Mbd2^−/−) groups. Our results demonstrated that deletion of Mbd2 in PyMT mice impedes primary tumor growth and lung metastasis at the experimental endpoint (postnatal week 20). Transcriptomic and proteomic analyses of primary tumors revealed that Mbd2 deletion abrogates the expression of several key determinants involved in epithelial-to-mesenchymal transition, such as neural cadherin (N-cadherin) and osteopontin. Importantly, loss of the Mbd2 gene impairs the activation of the PI3K/AKT pathway, which is required for PyMT-mediated oncogenic transformation, growth, and survival of breast tumor cells. Taken together, the results of this study provide a rationale for further development of epigenetic therapies targeting Mbd2 to inhibit the progression of breast cancer.

Breast cancer (BCa) is the most prevalent cancer in females and has a high rate of mortality, especially due to increased metastasis to skeletal and non-skeletal sites. Despite the marked clinical accomplishment of immune checkpoint inhibitor (CPI) therapy in patients with several cancers, it has had limited success in luminal subtypes of BCa. Accordingly, recent efforts have focused on combination therapy with CPI, including epigenetic modulators, to increase response rates of CPI in luminal BCa. We have previously shown that S-adenosylmethionine (SAM), the ubiquitous methyl donor, has strong anti-cancer effects in various cancers, including all subtypes of BCa. In the current study, we took a novel approach and examined the effect of CPI alone and in combination with SAM on tumor growth and metastasis in a syngeneic mouse model of luminal B BCa. We showed that SAM decreases cell proliferation, colony-formation (survival), and invasion of luminal B BCa cell lines (Eo771, R221A) in vitro. In in vivo studies, in Eo771 tumor-bearing mice, either SAM or anti-PD-1 antibody treatment alone significantly reduced tumor growth and progression, while the SAM+anti-PD-1 combination treatment had the highest anti-cancer efficacy of all groups. The SAM+anti-PD-1 combination reduced the percentage of animals with lung metastasis, as well as total metastatic lesion area, compared to control. Additionally, the SAM+anti-PD-1 combination significantly reduced the skeletal lesion area and protected tibial integrity to a greater extent than the monotherapies in an Eo771 bone metastasis model. Transcriptome analysis of Eo771 primary tumors revealed significant downregulation of pro-metastatic genes, including Matrix metalloproteinases (MMPs) and related pathways. On the other hand, CD8⁺ T cell infiltration, CD8⁺ T cell cytotoxicity (elevated granzymes), and immunostimulatory genes and pathways were significantly upregulated by the combination treatment. The results presented point to a combination of SAM with CPI as a possible treatment for luminal B BCa that should be tested in clinical studies.

Vitamin D regulates a plethora of physiological processes in the human body and has been proposed to exert several anticancer effects. Epigenetics plays an important role in regulating vitamin D ac... Graphical abstract Plain language summary Vitamin D regulates various physiological processes in the body and could have anticancer effects. These anticancer effects are the result of interactions between many factor... Tweetable abstract The present review describes the role of Vitamin D and epigenetic factors in tumorigenesis and their roles as potential targets for #ColorectalCancer treatment.

Fibrinolysis is a crucial physiological process that helps to maintain a hemostatic balance by counteracting excessive thrombosis. The components of the fibrinolytic system are well established and are associated with a wide array of physiological and pathophysiological processes. The aberrant expression of several components, especially urokinase-type plasminogen activator (uPA), its cognate receptor uPAR, and plasminogen activator inhibitor-1 (PAI-1), has shown a direct correlation with increased tumor growth, invasiveness, and metastasis. As a result, targeting the fibrinolytic system has been of great interest in the field of cancer biology. Even though there is a plethora of encouraging preclinical evidence on the potential therapeutic benefits of targeting the key oncogenic components of the fibrinolytic system, none of them made it from “bench to bedside” due to a limited number of clinical trials on them. This review summarizes our existing understanding of the various diagnostic and therapeutic strategies targeting the fibrinolytic system during cancer.

DNA and RNA methylation play a vital role in the transcriptional regulation of various cell types including the differentiation and function of immune cells involved in pro- and anti-cancer immunity. Interactions of tumor and immune cells in the tumor microenvironment (TME) are complex. TME shapes the fate of tumors by modulating the dynamic DNA (and RNA) methylation patterns of these immune cells to alter their differentiation into pro-cancer (e.g., regulatory T cells) or anti-cancer (e.g., CD8+ T cells) cell types. This review considers the role of DNA and RNA methylation in myeloid and lymphoid cells in the activation, differentiation, and function that control the innate and adaptive immune responses in cancer and non-cancer contexts. Understanding the complex transcriptional regulation modulating differentiation and function of immune cells can help identify and validate therapeutic targets aimed at targeting DNA and RNA methylation to reduce cancer-associated morbidity and mortality.