Prostate cancer (PCa) is one of the most common cancers in European males. Although therapeutic approaches have changed in recent years, and several new drugs have been approved by the Food and Drug Administration (FDA), androgen deprivation therapy (ADT) remains the standard of care. Currently, PCa represents a clinical and economic burden due to the development of resistance to ADT, paving the way to cancer progression, metastasis, and to long-term side effects induced by ADT and radio-chemotherapeutic regimens. In light of this, a growing number of studies are focusing on the tumor microenvironment (TME) because of its role in supporting tumor growth. Cancer-associated fibroblasts (CAFs) have a central function in the TME because they communicate with prostate cancer cells, altering their metabolism and sensitivity to drugs; hence, targeted therapy against the TME, and, in particular, CAFs, could represent an alternative therapeutic approach to defeat therapy resistance in PCa. In this review, we focus on different CAF origins, subsets, and functions to highlight their potential in future therapeutic strategies for prostate cancer.

Neuropathic pain (NP) is a chronic condition resulting from damaged pain-signaling pathways. It is a debilitating disorder that affects up to 10% of the world’s population. Although opioid analgesics are effective in reducing pain, they present severe risks; so, there is a pressing need for non-opioid pain-relieving drugs. One potential alternative is represented by sigma-1 receptor (S1R) antagonists due to their promising analgesic effects. Here, we report the synthesis and biological evaluation of a series of S1R antagonists based on a 2-aryl-4-aminobutanol scaffold. After assessing affinity toward the S1R and selectivity over the sigma-2 receptor (S2R), we evaluated the agonist/antagonist profile of the compounds by investigating their effects on nerve growth factor-induced neurite outgrowth and aquaporin-mediated water permeability in the presence and absence of oxidative stress. (R/S)-RC-752 emerged as the most interesting compound for S1R affinity (K_i S1R = 6.2 ± 0.9) and functional antagonist activity. Furthermore, it showed no cytotoxic effect in two normal human cell lines or in an in vivo zebrafish model and was stable after incubation in mouse plasma. (R/S)-RC-752 was then evaluated in two animal models of NP: the formalin test and the spinal nerve ligation model. The results clearly demonstrated that compound (R/S)-RC-752 effectively alleviated pain in both animal models, thus providing the proof of concept of its efficacy as an antinociceptive agent.

Different pathological conditions, including viral infections and cancer, can have a massive impact on the endoplasmic reticulum (ER), causing severe damage to the cell and exacerbating the disease. In particular, coronavirus infections, including SARS coronavirus-2 (SARS-CoV-2), responsible for COVID-19, cause ER stress as a consequence of the enormous amounts of viral glycoproteins synthesized, the perturbation of ER homeostasis and the modification of ER membranes. Therefore, ER has a central role in the viral life cycle, thus representing one of the Achilles’ heels on which to focus therapeutic intervention. On the other hand, prolonged ER stress has been demonstrated to promote many pro-tumoral attributes in cancer cells, having a key role in tumor growth, metastasis and response to therapies. In this report, adopting a repurposing approach of approved drugs, we identified the antiplatelet agent ticlopidine as an interferent of the unfolded protein response (UPR) via sigma receptors (SRs) modulation. The promising results obtained suggest the potential use of ticlopidine to counteract ER stress induced by viral infections, such as COVID-19, and cancer.

Spheroids, a part of 3D cell culture systems, are crucial models for bridging in-vitro and in-human studies. However, achieving reliable standardization remains difficult, even when comparing spheroids of similar diameters. The challenge arises due to their cross-sectional architecture, which increases heterogeneity and affects biological outcomes. Here we present a novel solution that integrates Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA) with Biophysical Characterization (PCA-BC). This approach allows for the identification and classification of variability within and across spheroid populations, offering insights into factors that contribute to heterogeneity. Additionally, it highlights the impact of different operators on spheroid development. The PCA-BC method enables real-time analysis of spheroid samples, facilitating the identification of variability across 3D populations. The integration of PCA and HCA with biophysical characterization provides a clear and efficient means to monitor sample heterogeneity. It also helps track how different operators influence the results, improving overall standardization in 3D cell cultures. By offering structural insights into spheroid heterogeneity, the PCA-BC approach supports more informed decision-making. This significantly improves workflow efficiency, conserving both time and resources, and enhances the reliability of 3D cell culture experiments.

RC-106 is a Sigma receptor modulator under investigation as a potential therapeutic agent for glioblastoma (GBM). Here, we report a multigram-scale synthesis based on green chemistry principles, its comprehensive solid-state characterization, and its preclinical evaluation in vitro and in vivo. ADMET prediction indicated good drug-likeness, absence of PAINS features, compliance with Lipinski criteria, favorable metabolic stability, and the ability to cross the blood-brain barrier. The optimized synthetic process to obtain RC-106·HCl (>99% purity) for solid-state characterization performed by TGA, DSC, FT-IR, XRPD, and SEM, confirming the compound crystalline nature and appropriate thermal behavior. in vivo, RC-106 consistently reduced cell viability in both commercial (U87, A172) and patient-derived GBM cell lines (G34, G48) under hypoxic conditions, inducing apoptosis and neurosphere disaggregation with IC₅₀ values between 44 and 54 μM. In vivo, using a patient-derived orthotopic xenograft (PDOX) mouse model, daily administration of RC-106·HCl (20 mg/kg) was well tolerated, with no observable systemic or neurological toxicity, as evidenced by stable body weight and preserved motor performance. Intraperitoneal administration of RC-106·HCl, although it did not definitively arrest tumor growth, resulted in a time-dependent reduction in intracranial GBM burden in our patient-derived orthotopic xenograft (PDOX) model. Pharmacokinetic and biodistribution analyses supported rapid absorption and distribution to the brain and spinal cord. Overall, RC-106·HCl shows promising in vitro anti-GBM activity and an excellent safety profile in vivo, while further optimization is needed to enhance its pharmacokinetic properties and therapeutic effectiveness.

Three-dimensional (3D) cultures represent an advanced approach for mimicking the architecture and cellular complexity of human tissues, providing a valuable alternative to conventional two-dimensional models. In particular, 3D bioprinting enables the generation of precise, standardized and biomimetic tissue constructs, with controlled spatial organization. The aim of the study was to design a novel 3D bioprinted full-thickness human skin model 3D protocol able to replicate human skin layers. The model is based on a high-concentration type I collagen bioink combined with dermal fibroblasts and keratinocytes, in which the use of dense collagen I enhances the mechanical properties and biomimetic features of the construct, providing dermal and epidermal contributions, and a physiologically relevant extracellular matrix. The skin like tissue (SLT) obtained was evaluated using established irritant and sensitizer controls, demonstrating its ability to respond to chemically induced stimuli without implying quantitative performance comparisons. Overall, this 3D skin model provides a biologically relevant and reproducible platform for early-stage testing of topically applied compounds, with potential applications in cosmetic safety assessment, drug screening, and alternative in vitro testing strategies.