Cesarina Giallongo

Background/Objectives: Radiotherapy (RT) is a fundamental treatment for brain tumors but often leads to long-term neurotoxicity, partly caused by microglial dysfunction. In this study, we used long-read RNA sequencing (LR-RNA-seq) to map the isoform-specific transcriptome of human microglial HMC3 cells after RT exposure. Methods: The HMC3 human microglial cell line was treated with standard radiotherapy and, after RNA extraction and library preparation, LR-RNA-seq was performed. Further bioinformatic analyses allowed to investigate on the isoform switching pattern after radiotherapy exposure. Results: Gene-level analysis found 591 differentially expressed genes, with positive enrichment of PI3K/AKT-related pathways and negative enrichment of translation-associated processes. Isoform-level analysis uncovered 827 significant isoform switching events across 420 genes, often involving shifts from protein-coding transcripts to non-coding variants. Functional annotation showed that most isoform switches resulted in the loss of open reading frames and protein domains, especially in ribosomal proteins and translation machinery components. Exploratory proteomic analysis identified 191 differentially abundant proteins, including a significant reduction in RPL7A, providing partial protein-level support for translation-associated transcriptomic remodeling. Conclusions: RT-induced stress causes qualitative transcriptomic remodeling beyond changes in overall gene expression. These findings indicate that radiotherapy induces coordinated gene-, transcript-, and isoform-level changes in HMC3 cells. The alterations observed in translation-associated genes, together with partial proteomic concordance, identify candidate mechanisms for future functional investigation.

Hepcidin, a 25-amino-acid peptide hormone produced primarily by hepatocytes, is the master regulator of systemic iron homeostasis. By binding the cellular iron exporter ferroportin and inducing its internalization and lysosomal degradation, hepcidin restricts iron entry into plasma from enterocytes, macrophages, and hepatocytes. Its transcription is governed by an intricate molecular network that integrates iron status, erythropoietic demand, oxygen tension, and inflammation, with the BMP–HJV–ALK2/SMAD axis acting as the canonical activating pathway and erythroferrone (ERFE) and matriptase-2 (TMPRSS6) as physiological suppressors. Dysregulation of hepcidin underpins a wide spectrum of human diseases: insufficient hepcidin drives hereditary hemochromatosis and the iron overload of congenital and acquired ineffective erythropoiesis diseases and other ineffective erythropoiesis syndromes, whereas excessive or inappropriate hepcidin contributes to anemia of inflammation, anemia of chronic kidney disease, iron-restricted erythropoiesis in cancer, the iron-restrictive anemia of myelofibrosis, and pathogen-restrictive nutritional immunity. Within the myeloproliferative neoplasm spectrum, the divergent hepcidin patterns observed in polycythemia vera (suppressed) and myelofibrosis (inappropriately elevated through dual BMP/ACVR1/SMAD and IL-6/STAT3 hyperactivation) exemplify the clinical relevance of this axis and underpin two opposite pharmacologic strategies. Over the past decade, hepcidin pathway pharmacology has matured from proof-of-concept to regulatory milestones, shifting perspectives on several diseases and markedly improving clinical approaches.

Ethanol-induced hepatocellular injury is characterized by redox imbalance and mitochondrial dysfunction, which critically determine cellular adaptation to metabolic stress. Rather than acting as simple antioxidants, effective nutraceutical strategies may enhance endogenous adaptive responses that preserve cellular homeostasis. Pistachio (Pistacia vera L.) leaves, an underexplored agricultural by-product, represent a potential source of bioactive compounds with redox-modulating properties. In this study, we investigated the effects of pistachio leaf extract (PLE) in human HepG2 hepatocytes exposed to 1% ethanol (EtOH). EtOH reduced cell viability and increased intracellular and mitochondrial reactive oxygen species (ROS), while triggering compensatory responses including glutathione accumulation and activation of metabolic and inflammatory pathways. PLE alone induced a mild cytosolic ROS signal and increased PGC-1α expression, consistent with transcriptional changes potentially associated with hormetic activation of mitochondrial regulatory pathways. Notably, co-treatment with EtOH and PLE significantly reduced both cytosolic and mitochondrial ROS levels, prevented excessive glutathione accumulation and was associated with transcriptional changes consistent with improved glutathione-related adaptive responses. These effects were associated with transcriptional modulation of genes involved in mitochondrial biogenesis and metabolic adaptation, including PGC-1α, TFAM, SIRT1, PPARα, and UCP2. In parallel, PLE attenuated EtOH-induced IL-6 expression without preventing FAS upregulation, indicating a selective dissociation between inflammatory and lipogenic responses. Overall, our findings suggest that PLE may act as a modulator of redox-dependent signaling rather than as a direct antioxidant and is associated with transcriptional and functional signatures consistent with mitochondrial adaptive responses to ethanol-induced stress. These results support the potential of pistachio leaf-derived phytocomplexes as nutraceutical modulators of hepatocellular adaptive responses.

Supplementary Figure S5. Gene set enrichment analysis in MM cells following ND-646 treatment, and evaluation of cell growth under combined treatment with ND-646 and simvastatin.

Supplementary Figure S4. Impact of pharmacological inhibition (ND-630 and ND-646) and ACC1 knockdown on MM cell viability, proliferation, and de novo lipogenesis (DNL) rates in MYC overexpressed MM cells.

Supplementary Figure S1. Expression profiling and correlation analysis of MYC and ACC1 in patients with newly diagnosed multiple myeloma (NDMM).

Supplementary Figure S2. Quantitative analysis of ACACA expression in multiple myeloma (MM) cell lines.

Supplementary Figure S3. Representative western blot analysis showing protein levels of ACC1, phosphorylated ACC (pACC), and MYC in MM cells.

Purpose: In multiple myeloma, tumor cells reprogram metabolic pathways to sustain growth and monoclonal immunoglobulin production. This study examines acetyl-CoA carboxylase 1 (ACC1), the enzyme driving the rate-limiting step in de novo lipogenesis, in multiple myeloma metabolic reprogramming, particularly in c-MYC (MYC)–driven subtypes. Experimental Design: ACC1 expression was evaluated across multiple myeloma genetic subgroups, focusing on MYC translocations. Functional studies using ACC1 inhibitors and genetic knockdown assessed multiple myeloma cell growth, lipid synthesis, and metabolic homeostasis in vitro and in vivo. The role of MYC overexpression in ACC1 sensitivity was examined, with palmitate rescue experiments. Lipidomic analysis and assessments of endoplasmic reticulum (ER) stress, protein translation, and oxidative damage elucidated underlying mechanisms. Results: ACC1 was overexpressed in MYC-translocated multiple myeloma. Its inhibition or knockdown reduced multiple myeloma cell growth in vitro and in vivo, particularly in MYC-overexpressing cells. ACC1 knockdown suppressed de novo lipid synthesis, partially rescued by palmitate. Lipidomic disruptions increased cholesterol ester desaturation and altered phospholipid ratios, inducing ER stress, impaired translation, protein carbonylation, oxidative damage, and apoptosis. Conclusions: ACC1 is a metabolic vulnerability in MYC-driven multiple myeloma. Inhibiting ACC1 disrupts lipid homeostasis, induces ER stress, and causes oxidative damage, impairing cell survival. Targeting lipid synthesis pathways, especially in MYC-dependent subtypes, offers a promising therapeutic strategy for multiple myeloma.

Background: Immune system impairment is frequently reported in patients affected by hemoglobinopathies due to various mechanisms, including iron accumulation, antigenic stimulation due to numerous transfusions, chronic hemolysis, and a general hyperinflammatory state. For these reasons, the antigenic immune response after a vaccine risks being ineffective. Methods: We evaluated the anti-spike IgG production after two doses of vaccine for SARS-CoV-2 in patients affected by hemoglobinopathies. Results: All 114 enrolled patients (100%) developed adequate antibody production, with a median value of serum IgG of 2184.4 BAU/mL (IQR 1127.4–3502.9). The amount of antibody was unrelated to any other clinical characteristics evaluated, including transfusion dependence or non-transfusion dependence, age, gender, disease type, ferritin, blood count, spleen status, and therapy with hydroxyurea or iron chelators (in all the cases p > 0.05). Moreover, 47 (41.2%) patients developed breakthrough SARS-CoV-2 infection during the first 2 years of follow-up after vaccination, all with a mildly symptomatic course, without requiring hospitalization or experiencing a significative drop in hemoglobin values, allowing for a slight delay in their transfusion regimen. Conclusion: Vaccination against COVID-19 is safe and effective for patients affected by hemoglobinopathies, ensuring adequate protection from severe infection.