Serine/threonine‐protein kinase B‐raf (BRAF) mutations are found in 8–15% of colorectal cancer patients and identify a subset of tumors with poor outcome in the metastatic setting. We have previously reported that BRAF‐mutant human cells display a high rate of protein production, causing proteotoxic stress, and are selectively sensitive to the proteasome inhibitors bortezomib and carfilzomib. In this work, we tested whether carfilzomib could restrain the growth of BRAF‐mutant colorectal tumors not only by targeting cancer cells directly, but also by promoting an immune‐mediated antitumor response. In human and mouse colorectal cancer cells, carfilzomib triggered robust endoplasmic reticulum stress and autophagy, followed by the emission of immunogenic‐damage‐associated molecules. Intravenous administration of carfilzomib delayed the growth of BRAF‐mutant murine tumors and mobilized the danger‐signal proteins calreticulin and high mobility group box 1 (HMGB1). Analyses of drug‐treated samples revealed increased intratumor recruitment of activated cytotoxic T cells and natural killers, concomitant with the downregulation of forkhead box protein P3 (Foxp3)⁺ T‐cell surface glycoprotein CD4 (CD4)⁺ T cells, indicating that carfilzomib promotes reshaping of the immune microenvironment of BRAF‐mutant murine colorectal tumors. These results will inform the design of clinical trials in BRAF‐mutant colorectal cancer patients.

Background: O-6-methylguanine-DNA methyltransferase is responsible for the direct repair of O6-methylguanine lesions induced by alkylating agents, including temozolomide. O-6-methylguanine-DNA methyltransferase promoter hypermethylation is a well-established biomarker for temozolomide response in glioblastoma patients, also correlated with therapeutic response in colorectal cancer. Objectives: The ARETHUSA clinical trial aims to stratify colorectal cancer patients based on their mismatch repair status. Mismatch repair-deficient patients are eligible for treatment with immune checkpoint inhibitors (anti-PDL-1), whereas mismatch repair-proficient samples are screened for O-6-methylguanine-DNA methyltransferase promoter methylation to identify those suitable for temozolomide treatment. Methods: In this context, a subset of ARETHUSA metastatic colorectal cancer samples was used to compare two different techniques for assessing O-6-methylguanine-DNA methyltransferase hypermethylation: Methyl-BEAMing, a highly sensitive digital PCR approach that combines emulsion PCR and flow cytometry, and droplet digital PCR, a more automated procedure that enables the rapid, operator-independent analysis of a large number of samples. Results: Our study clearly demonstrates that the results obtained using Methyl-BEAMing and droplet digital PCR are comparable, with both techniques showing similar accuracy, sensitivity, and reproducibility. Conclusions: Digital droplet PCR proved to be an efficient method for detecting gene promoter methylation. However, the Methyl-BEAMing method has proved more sensitive for detecting low quantities of DNA.

Osteoarthritis (OA) is one of the most common joint diseases worldwide, predominantly present in elderly people. Being a major source of pain for patients, it is debilitating and leads inevitably to a reduction in quality of life. The management of OA needs a personalized and multidimensional approach, resulting in the emergence of new regenerative and non-invasive methods, such as the use of micrografts. In this pilot study, Rigenera^® Technology was employed to obtain micrografts of cartilage tissue to be injected into the knees of 10 patients with osteoarthritic pain. To assess the efficacy of the treatment concerning pain reduction at this site, patients were asked to complete KOOS and WOMAC questionnaire and a VAS test before and after the procedure. The results presented in this article show how Rigenera^® treatment can potentially improve OA symptoms, alleviating pain in patients.

High-dose vitamin C (vitC) has shown superior anticancer activity in immunocompetent compared with immunodeficient mice, suggesting an immunodependent mechanism of action. Because high-dose vitC generates substantial reactive oxygen species (ROS), we hypothesized that ROS could induce immunogenic cell death (ICD), a regulated form of cell death characterized by the release of damage-associated molecular patterns (DAMPs) that enhance antigen presentation and promote antitumor immunity. Spatial transcriptomic and immunofluorescence analyses were performed on breast and colon tumors from mice treated with high-dose vitC to quantify oxidative stress, DAMP mobilization, and immune cell infiltration. To dissect the functional role of ROS and ICD, mice were treated with high-dose vitC in combination with the antioxidant N-acetylcysteine or an antibody blocking calreticulin (an early DAMP essential for ICD induction). VitC treatment caused marked oxidative stress in tumors from both immunocompetent and immunocompromised mice; however, tumor growth inhibition occurred exclusively in immunocompetent animals. High-dose vitC triggered ROS-dependent exposure and release of the DAMPs calreticulin and HMGB1, respectively. This was accompanied by substantial remodeling of the tumor microenvironment, including increased infiltration of cytotoxic CD8⁺ T cells and natural killer cells, and a reduction in immunosuppressive regulatory T cells. Blocking calreticulin effectively disrupted the ICD cascade, prevented immune microenvironment remodeling, and completely abrogated the antitumor efficacy of vitC. Collectively, our study indicates that high-dose vitC promotes ICD by mobilizing calreticulin and HMGB1, and that calreticulin is a determinant of its immune-mediated antitumor activity. Implications: vitC induces ICD, supporting its use as an adjuvant to anticancer immunotherapy.

Background: Stretch marks or striae distensae (SD) are common dermal lesions caused by the disruption of collagen and elastin fibers in the skin, often triggered by rapid mechanical stretching. Despite the availability of numerous treatment modalities, from topical agents to energy‐based devices, no single therapy has demonstrated consistent, long‐term efficacy across all patient populations. The pathophysiology of SD involves complex alterations in the extracellular matrix (ECM), particularly affecting fibroblast activity and collagen/elastin synthesis.Aims: This pilot study aims to evaluate the clinical and molecular efficacy of autologous micrografting technology as a novel therapeutic option for SD. Specifically, it investigates the treatment's impact on ECM‐related gene expression and overall skin appearance.Patients/Methods: Fourteen patients (13 females, 1 male) with clinically evident SD were enrolled. All participants underwent a standardized treatment protocol comprising microneedling followed by intradermal injection of autologous micrografts, obtained via a minimally invasive procedure. Clinical assessments were performed through standardized photography at baseline, 1 month, and 6 months post‐treatment. In vitro assays were conducted on cultured human dermal fibroblasts exposed to the micrograft suspension.Results: Clinical evaluations showed noticeable aesthetic improvements, including reduced striae visibility and improved skin texture, with high patient‐reported satisfaction. Molecular analyses revealed the upregulation of key ECM genes, including COL4A1, COL6A1, and ELN, indicating enhanced fibroblast activation and regenerative potential.Conclusions: Autologous micrografting appears to be a promising, biologically active approach for SD treatment. It promotes ECM remodeling by stimulating fibroblast function and may represent a valuable addition to the therapeutic landscape for stretch marks.