Cancer Research, Aging, Biochemistry, Genetics and Molecular Biology, Cell Biology
39
Scopus Publications
Scopus Publications
Metabolic traits shape responses to LSD1-directed therapy in glioblastoma tumor-initiating cells Giulia Marotta, Daniela Osti, Elena Zaccheroni, Brunella Costanza, Stefania Faletti, et al. Science Advances, 2025 Lysine-specific histone demethylase 1A (LSD1) is an epigenetic regulator involved in various biological processes, including metabolic pathways. We demonstrated the therapeutic potential of its pharmacological inhibition in glioblastoma using DDP_38003 (LSD1i), which selectively targets tumor-initiating cells (TICs) by hampering their adaptability to stress. Through biological, metabolic, and omic approaches, we now show that LSD1i acts as an endoplasmic reticulum (ER) stressor, activating the integrated stress response and altering mitochondrial structure and function. These effects impair TICs’ oxidative metabolism and generate reactive oxygen species, further amplifying cellular stress. LSD1i also impairs TICs’ glycolytic activity, causing their metabolic decline. TICs with enhanced glycolysis benefit from LSD1-directed therapy. Conversely, metabolically silent TICs mantain ER and mitochondrial homeostasis, adapting to stress conditions, including LSD1i treatment. A dropout short hairpin RNA screening identifies postglycosylphosphatidylinositol attachment to proteins inositol deacylase 1 (PGAP1) as a mediator of resistance to LSD1i. Disruptions in ER and mitochondrial balance holds promise for improving LSD1-targeted therapy efficacy and overcoming treatment resistance.
High-Fat Diet Promotes Acute Promyelocytic Leukemia through PPARd-Enhanced Self-renewal of Preleukemic Progenitors Luca Mazzarella, Paolo Falvo, Marta Adinolfi, Giulia Tini, Elena Gatti, et al. Cancer Prevention Research, 2024 Risk and outcome of acute promyelocytic leukemia (APL) are particularly worsened in obese-overweight individuals, but the underlying molecular mechanism is unknown. In established mouse APL models (Ctsg-PML::RARA), we confirmed that obesity induced by high-fat diet (HFD) enhances leukemogenesis by increasing penetrance and shortening latency, providing an ideal model to investigate obesity-induced molecular events in the preleukemic phase. Surprisingly, despite increasing DNA damage in hematopoietic stem cells (HSC), HFD only minimally increased mutational load, with no relevant impact on known cancer-driving genes. HFD expanded and enhanced self-renewal of hematopoietic progenitor cells (HPC), with concomitant reduction in long-term HSCs. Importantly, linoleic acid, abundant in HFD, fully recapitulates the effect of HFD on the self-renewal of PML::RARA HPCs through activation of peroxisome proliferator-activated receptor delta, a central regulator of fatty acid metabolism. Our findings inform dietary/pharmacologic interventions to counteract obesity-associated cancers and suggest that nongenetic factors play a key role. Prevention Relevance: Our work informs interventions aimed at counteracting the cancer-promoting effect of obesity. On the basis of our study, individuals with a history of chronic obesity may still significantly reduce their risk by switching to a healthier lifestyle, a concept supported by evidence in solid tumors but not yet in hematologic malignancies. See related Spotlight, p. 47
A Rare Subset of Primary Tumor Cells with Concomitant Hyperactivation of Extracellular Matrix Remodeling and dsRNA-IFN1 Signaling Metastasizes in Breast Cancer Niccolò Roda, Andrea Cossa, Roman Hillje, Andrea Tirelli, Federica Ruscitto, et al. Cancer Research, 2023 Metastatic breast cancer has a poor prognosis and is largely considered incurable. A better understanding of the molecular determinants of breast cancer metastasis could facilitate development of improved prevention and treatment strategies. We used lentiviral barcoding coupled to single-cell RNA sequencing to trace clonal and transcriptional evolution during breast cancer metastasis and showed that metastases derive from rare prometastatic clones that are underrepresented in primary tumors. Both low clonal fitness and high metastatic potential were independent of clonal origin. Differential expression and classification analyses revealed that the prometastatic phenotype was acquired by rare cells characterized by the concomitant hyperactivation of extracellular matrix remodeling and dsRNA-IFN signaling pathways. Notably, genetic silencing of key genes in these pathways (KCNQ1OT1 or IFI6, respectively) significantly impaired migration in vitro and metastasis in vivo, with marginal effects on cell proliferation and tumor growth. Gene expression signatures derived from the identified prometastatic genes predict metastatic progression in patients with breast cancer, independently of known prognostic factors. This study elucidates previously unknown mechanisms of breast cancer metastasis and provides prognostic predictors and therapeutic targets for metastasis prevention. Significance: Transcriptional lineage tracing coupled with single-cell transcriptomics defined the transcriptional programs underlying metastatic progression in breast cancer, identifying prognostic signatures and prevention strategies.
Beyond Genetics: Metastasis as an Adaptive Response in Breast Cancer Federica Ruscitto, Niccolò Roda, Chiara Priami, Enrica Migliaccio, Pier Giuseppe Pelicci International Journal of Molecular Sciences, 2022 Metastatic disease represents the primary cause of breast cancer (BC) mortality, yet it is still one of the most enigmatic processes in the biology of this tumor. Metastatic progression includes distinct phases: invasion, intravasation, hematogenous dissemination, extravasation and seeding at distant sites, micro-metastasis formation and metastatic outgrowth. Whole-genome sequencing analyses of primary BC and metastases revealed that BC metastatization is a non-genetically selected trait, rather the result of transcriptional and metabolic adaptation to the unfavorable microenvironmental conditions which cancer cells are exposed to (e.g., hypoxia, low nutrients, endoplasmic reticulum stress and chemotherapy administration). In this regard, the latest multi-omics analyses unveiled intra-tumor phenotypic heterogeneity, which determines the polyclonal nature of breast tumors and constitutes a challenge for clinicians, correlating with patient poor prognosis. The present work reviews BC classification and epidemiology, focusing on the impact of metastatic disease on patient prognosis and survival, while describing general principles and current in vitro/in vivo models of the BC metastatic cascade. The authors address here both genetic and phenotypic intrinsic heterogeneity of breast tumors, reporting the latest studies that support the role of the latter in metastatic spreading. Finally, the review illustrates the mechanisms underlying adaptive stress responses during BC metastatic progression.
P66SHC deletion improves fertility and progeric phenotype of late-generation TERC-deficient mice but not their short lifespan Marco Giorgio, Massimo Stendardo, Enrica Migliaccio, Pier Giuseppe Pelicci Aging Cell, 2016 Oxidative stress and telomere attrition are considered the driving factors of aging. As oxidative damage to telomeric DNA favors the erosion of chromosome ends and, in turn, telomere shortening increases the sensitivity to pro‐oxidants, these two factors may trigger a detrimental vicious cycle. To check whether limiting oxidative stress slows down telomere shortening and related progeria, we have investigated the effect of p66SHC deletion, which has been shown to reduce oxidative stress and mitochondrial apoptosis, on late‐generation TERC (telomerase RNA component)‐deficient mice having short telomeres and reduced lifespan. Double mutant (TERC−/− p66SHC−/−) mice were generated, and their telomere length, fertility, and lifespan investigated in different generations. Results revealed that p66SHC deletion partially rescues sterility and weight loss, as well as organ atrophy, of TERC‐deficient mice, but not their short lifespan and telomere erosion.
Modelling the p53/p66Shc aging pathway in the shortest living vertebrate Nothobranchius furzeri Chiara Priami, Giulia De Michele, Franco Cotelli, Alessandro Cellerino, Marco Giorgio, et al. Aging and Disease, 2015 Oxidative stress induced by reactive oxygen species (ROS) increases during lifespan and is involved in aging processes. The p66Shc adaptor protein is a master regulator of oxidative stress response in mammals. Ablation of p66Shc enhances oxidative stress resistance both in vitro and in vivo. Most importantly, it has been demonstrated that its deletion retards aging in mice. Recently, new insights in the molecular mechanisms involving p66Shc and the p53 tumor suppressor genes were given: a specific p66Shc/p53 transcriptional regulation pathway was uncovered as determinant in oxidative stress response and, likely, in aging. p53, in a p66Shc-dependent manner, negatively downregulates the expression of 200 genes which are involved in the G2/M transition of mitotic cell cycle and are downregulated during physiological aging. p66Shc modulates the response of p53 by activating a p53 isoform (p44/p53, also named Delta40p53). Based on these latest results, several developments are expected in the future, as the generation of animal models to study aging and the evaluation of the use of the p53/p66Shc target genes as biomarkers in aging related diseases. The aim of this review is to investigate the conservation of the p66Shc and p53 role in oxidative stress between fish and mammals. We propose to approach this study trough a new model organism, the annual fish Nothobranchius furzeri, that has been demonstrated to develop typical signs of aging, like in mammals, including senescence, neurodegeneration, metabolic disorders and cancer.
The influence of Shc proteins on life span in mice Jon J. Ramsey, Dianna Tran, Marco Giorgio, Stephen M. Griffey, Amanda Koehne, et al. Journals of Gerontology Series A Biological Sciences and Medical Sciences, 2014 The signaling molecule p66Shc is often described as a longevity protein. This conclusion is based on a single life span study that used a small number of mice. The purpose of the present studies was to measure life span in a sufficient number of mice to determine if longevity is altered in mice with decreased Shc levels (ShcKO). Studies were completed at UC Davis and the European Institute of Oncology (EIO). At UC Davis, male C57BL/6J WT and ShcKO mice were fed 5% or 40% calorie-restricted (CR) diets. In the 5% CR group, there was no difference in survival curves between genotypes. There was also no difference between genotypes in prevalence of neoplasms or other measures of end-of-life pathology. At 40% calorie restriction group, 70th percentile survival was increased in ShcKO, while there were no differences between genotypes in median or subsequent life span measures. At EIO, there was no increase in life span in ShcKO male or female mice on C57BL/6J, 129Sv, or hybrid C57BL/6J-129Sv backgrounds. These studies indicate that p66Shc is not a longevity protein. However, additional studies are needed to determine the extent to which Shc proteins may influence the onset and severity of specific age-related diseases.
Deletion of p66shc in mice increases the frequency of size-change mutations in the lacZ transgene Elena Beltrami, Antonella Ruggiero, Rita Busuttil, Enrica Migliaccio, Pier Giuseppe Pelicci, et al. Aging Cell, 2013 SummaryUpon oxidative challenge the genome accumulates adducts and breaks that activate the DNA damage response to repair, arrest, or eliminate the damaged cell. Thus, reactive oxygen species (ROS) generated by endogenous oxygen metabolism are thought to affect mutation frequency. However, few studies determined the mutation frequency when oxidative stress is reduced. To test whether in vivo spontaneous mutation frequency is altered in mice with reduced oxidative stress and cell death rate, we crossed p66Shc knockout (p66KO) mice, characterized by reduced intracellular concentration of ROS and by impaired apoptosis, with a transgenic line harboring multiple copies of the lacZ mutation reporter gene as part of a plasmid that can be recovered from organs into Escherichia coli to measure mutation rate. Liver and small intestine from 2‐ to 24‐month‐old, lacZ (p66Shc+/+) and lacZp66KO mice, were investigated revealing no difference in overall mutation frequency but a significant increase in the frequency of size‐change mutations in the intestine of lacZp66KO mice. This difference was further increased upon irradiation of mice with X‐ray. In addition, we found that knocking down cyclophilin D, a gene that facilitates mitochondrial apoptosis acting downstream of p66Shc, increased the size‐change mutation frequency in small intestine. Size‐change mutations also accumulated in death‐resistant embryonic fibroblasts from lacZp66KO mice treated with H2O2. These results indicate that p66Shc plays a role in the accumulation of DNA rearrangements and suggest that p66Shc functions to clear damaged cells rather than affect DNA metabolism.
P53 and aging: Role of p66Shc Enrica Migliaccio, Marco Giorgio, Pier Giuseppe Pelicci Aging, 2013 The p53 protein was discovered many years ago as a tumour suppressor gene and despite the wealth of information that has accumulated, a complete understanding of how p53 functions remains still elusive. p53 is a key regulator of the checkpoint response to DNA damage in mammal cells and is affected by loss-of-function mutations in the majority of human cancer. In various normal adult tissues, basal levels of p53 protein are low, but specific stress signals induce post-translation modifications and its stabilization, leading to activation of distinct transcriptional programmers. In response to stress signals p53 mainly acts to repair DNA damage and depending on the severity of the damage, the cell microenvironment and the cell type, p53 can orchestrate different cellular outcomes such as apoptosis and senescence. In addition to its nuclear activity, p53 also possesses biological activities that are transcription-independent, indeed p53 functions also in the mitochondria and cytoplasm, where, depending on the kind of stress either triggers apoptosis or inhibits autophagy [1, 2]. Then, the DeltaNp53 isoform, that lacks the transactivation domain, contributes to regulate p53 functions [3]. Notably, the over-expression of DeltaNp53 (p44) in mice affects IGF-1 induced gene expression and accelerates aging [4]. Cells from these mice are more susceptible to both p53-mediated apoptosis and senescence, suggesting that enhanced tumor suppressive response comes at the cost of accelerated aging. Indeed, emerging evidence suggested that p53 and DNA damage checkpoints have also a prominent but still not well characterized role in the regulation of aging in worm, mice and human [5]. How p53 differentiate between different stresses so that its activation leads to the correct response and whether aging is part of the tumorsuppressive function of p53 are outstanding questions. Emerging findings suggest that p53 activity on specific target is regulated by specific p53 isoforms, in the context of specific activating-signals [3]. Typically, p53-dependent growth arrest in G1 phases of the cell cycle have been attributed to transcriptional activity of p53 [both activation and repression] but contrary to the p53-transcription activation the biological significance of p53-mediate transcription repression and its role in G2/M arrest of the cell cycle is still largely undisclosed. p44/p53 (p47 in human) is an N-terminally truncated p53 isoform, which forms homo-oligomers and hetero-oligomers with p53, and induces G2/M cell-cycle arrest in response to serum deprivation or endoplasmic reticulum (ER) stress [6]. However, the regulation and physiological role of this p44 isoform are just starting to become known and its pro-aging function remains the unique indication regarding its putative physiological role [4]. Recent studies have demonstrated the presence of internal ribosome entry site (IRES) sequences in p53 mRNA that mediated the translation of both full-length and p44/p53 isoforms and represent a novel control of p53 gene expression and activity [6]. Then, the p44 isoform translation was found boosted by ER stress and plays an essential role in the unfolded protein response (UPR) that leads to a p53-mediated G2 cell cycle arrest [6]. Recently, it has been demonstrated that p53-dependent G2 arrest following ER stress is mediated by the p44 isoform [6]. One intriguing open question on p53 role in life span determination comes from the assumption that the increase of p53 activity is associated at acceleration of aging and at decrease of cancer incidence, and in opposite way, the attenuation of its activity correlates with short life span and increase of incidence of cancer. However, contrary to this idea, it appeared that many of the long-lived strains models, including p66Shc knockout mice [7], attenuation of p53 function does not correlated with increased of cancer. Recent findings suggest also that p53 and p66Shc are genetically and functionally linked [8]. p66Shc is a vertebrate protein whose targeted deletion in mice induces resistance to a variety of aging-associated diseases, including obesity, atherosclerosis, ischemic injury and diabetes, and extends lifespan [7]. In mitochondria, p66Shc uses reducing equivalents of the electron-transfer chain via direct oxidation of cytochrome c to generate pro-apoptotic reactive oxygen species [ROS], acting as a stress-induced red-ox enzyme, that also function as signaling molecules to activate specific targets [7]. Our group demonstrates that the p53 transcriptional-response to oxidative stress involves a large number of G2/M-mitosis genes and depends on p66Shc. In particular, the expression of p66Shc is indispensable for the function of the p53 isoform (p44/p53) to induce of G2/M cell cycle arrest, after oxidative stress. Consistently, p66Shc deletion decreased some of the signs of premature aging observed in mice overexpressing the p44 isoform [8], such as, decrease lifespan, decrease of fertility, premature accumulation of fat tissue in the liver, premature thymic involution and alopecia, demonstrating that p66Shc is critical for the biological effects of p44/p53 also in vivo [8]. Based on these results we hypothesized the tumour suppressive and the pro-aging functions of p53 are managed by two distinct pathways (Figure (Figure1)1) and that p66Shc is a critical component of the aging pathway. This one is triggered by the accumulation of oxidation damaged products (mainly proteins) that induces UPR and activates a specific p53 pathway that depends on the p66Shc redox activity and the p44 short isoform. Figure 1 Distinct p53 pathways regulate tumour suppression and aging. Oxidative stress activates a specific specific transcriptional response, mediated by p66Shc and p44/p53 isoforms, which regulates cellular senescence and aging. As consequence, the existence of two different and not redundant p53 pathways explains why the abrogation of the p53 aging function, as it occurs in p66Shc null mice, does not increase tumour formation.
Selective expression and constitutive phosphorylation of Src-homology-2 and collagen-homology domains proteins in the CD34+ fraction of chronic myelogenous leukemias Cancer Research, 2000
Not all Shc's roads lead to Ras Laura Bonfini, Enrica Migliaccio, Giuliana Pelicci, Luisa Lanfrancone, PierGiuseppe Pelicci Trends in Biochemical Sciences, 1996
