The Prokineticin system is downregulated in idiopathic rapid eye movement sleep behavior disorder: evidence from olfactory neurons Piergiorgio Grillo, Daniela Maftei, Alessandra Calculli, Tommaso Schirinzi, Simone Mauramati, Martina Vincenzi, Maria Grazia Di Certo, Francesca Gabanella, Deborah Di Martino, Marco Benazzo, Cinzia Severini, Roberta Lattanzi, Antonio Pisani, Michele Terzaghi Sleep, 2026 Study Objectives PROK2 is a peptide expressed in the adult brain mediating neuroprotective functions. Previous studies reported an upregulation of prokineticin system in Parkinson’s disease (PD), but evidence in prodromal α-synucleinopathies was lacking. We investigated the expression of prokineticin-2 (PROK2) and its receptors (PKR1 and PKR2), along with oligomeric α-synuclein (oligo α-syn) as a marker of α-synuclein pathology, in olfactory neurons (ONs) from individuals with idiopathic rapid eye movement sleep behavior disorder (iRBD). Methods Olfactory neurons, obtained by nasal brush from 28 idiopathic rapid eye movement sleep behavior disorder subjects (age: 71.2 ± 7.4 years; males: 89.3%; duration: 4.9 ± 2.5 years) and 28 healthy controls (HCs) (age:67.2 ± 11.5 years; males:64.2%), were analyzed using real-time polymerase-chain-reaction (RT-PCR), immunofluorescence (IF), and western blot (WB). In a subgroup of subjects, results were validated in serum. Results In the idiopathic rapid eye movement sleep behavior disorder group, prokineticin-2 protein expression was reduced in both ONs (immunofluorescence: F(1,26) = 15.289, p < .001; western blot: F(1,12) = 9.073, p = .011) and serum compared with HCs (western blot: F(1,12) = 4.557, p = .050). Idiopathic rapid eye movement sleep behavior disorder subjects showed lower mRNA expression of prokineticin receptors compared with healthy controls (real-time polymerase-chain-reaction for prokineticin receptor-1: F(1,26) = 16.131, p < .001; real-time polymerase-chain-reaction for prokineticin receptor-2: F(1,39) = 4.946, p = .032). Oligo α-syn accumulation in olfactory neurons was higher in idiopathic rapid eye movement sleep behavior disorder than healthy controls, yet the difference only tended to statistical significance (immunofluorescence: F(1,18) = 3.169, p = .092). Conclusions In contrast with findings in Parkinson’s disease, we found a downregulation of prokineticin system in idiopathic rapid eye movement sleep behavior disorder. The causes of prokineticin system downregulation in this prodromal stage may be multiple. The absence of clear oligo α-syn accumulation, known trigger of prokineticin-2, may play a role. On the other hand, a lack of activation of this system might act as predisposing factor for the development of idiopathic rapid eye movement sleep behavior disorder and, subsequently, full-blown neurodegeneration.
miR-124 orchestrates epicardial-mesenchymal transformation and paracrine cardiomyocyte maturation in epicardial-specific Tcf21-PKR1 knockout mice Himanshu Arora, Martina Vincenzi, Anais Audebrand, Amin Kremic, Carmine Gentile, Laurent Desaubry, Canan G Nebigil Stem Cells, 2026 During the heart development, epicardial-to-mesenchymal transition (EMT) drives the production of progenitor cell populations that contribute to heart formation; however, the molecular control of EMT and its paracrine effects on cardiomyocytes remain poorly elucidated. Here, we defined a novel PKR1–miR-124–SNAI2 signaling axis that orchestrates EMT and coordinates myocardial maturation. Conditional deletion of the prokineticin receptor (PKR1) in mice Tcf21+ epicardial cells caused embryonic lethality and congenital heart disease-like anomalies, including ventricular rupture, arrhythmia, myocardial fibrosis, and impaired contractility. Transcriptomic profiling revealed marked upregulation of miR-124, concurrent with deregulation of EMT genes and signatures of immature cardiomyocytes. Mechanistically, miR-124 directly targets the 3′ untranslated region of SNAI2, suppressing this key EMT regulator, resulting in failed EMT, apoptosis, and fibrosis in the epicardium. Functional rescue through miR-124 inhibition or PKR1 reintroduction restores SNAI2 expression, revives EMT, enhances cell survival, and promotes proper cardiomyocyte maturation. Paracrine effects were substantiated by conditioned media and ex vivo assays, demonstrating that epicardial-derived miR-124 suppressed cardiomyocyte contractility and cardiac maturity gene expression—thereby functionally linking epicardial disruption to myocardial immaturity. These findings establish miR-124 as a critical mediator of epicardial–myocardial communication with PKR1 as its upstream regulator. By integrating epicardial plasticity, myocardial maturation, and ECM homeostasis, our work reveals that targeting the PKR1–miR-124–SNAI2 pathway offers a novel mechanistic framework and potential therapeutic target for preventing or treating congenital heart disease.
Identification of prokineticin-2 as potential pharmacodynamic biomarker for overcoming doxorubicin resistance in multicellular breast cancer spheroids Martina Vincenzi, Amin Kremic, Nathalie N. Tscheiller, Po‐Yen Hsu, Michael W. Y. Chan, Roberta Lattanzi, Canan G. Nebigil British Journal of Pharmacology, 2026 Background and purposeDespite advances in immunotherapy, doxorubicin (Dox) chemotherapy is still the irreplaceable first‐line therapy for solid tumours such as breast cancer. However, chemotherapy resistance is the major limiting factor, requiring the use of high doses of Dox to achieve the anti‐tumour actions, often leading to severe side effects. Unravelling the mechanisms behind chemoresistance and identifying potential biomarkers for mitigating this resistance could enhance current treatment strategies and improve patient outcomes.Experimental approachWe developed human 3D breast cancer spheroids (HBCSs) as a model that closely mimics in vivo tumour structure and microenvironment. Given that hypoxia and elevated levels of the angiogenic cytokine, prokineticin‐2 (PK2), are associated with chemoresistance to antiangiogenic therapy, we explored the effect of a hypoxia‐inducible factor (HIF‐1α) inhibitor on viability defect in HBCSs and the levels of PK2 in the conditioned medium following Dox treatment. We also assessed levels of HIF‐1α, active caspase‐3, TUNEL and reactive oxygen species (ROS), and CD73 enzymatic activity in HBCSs.Key resultsResults showed that HIF‐1α inhibitor increased viability defect in the Dox‐resistant HBCSs. Interestingly, at higher Dox concentrations, chemoresistance was mitigated independently of HIF‐1α and promoted apoptosis and ROS accumulation, which were correlated with PK2 release.Conclusions and implicationsOur findings provide the first evidence that PK2 may serve as a predictive pharmacodynamic marker, offering a potential strategy to overcome drug resistance in targeted cancer therapy.
Uncovering the role of prokineticin pathway on Epicardial Adipose Tissue (EAT) development and EAT-associated cardiomyopathy Martina Vincenzi, Canan G. Nebigil Trends in Cardiovascular Medicine, 2025 Epicardial adipose tissue (EAT), a unique fat depot surrounding the heart, plays a multifaceted role in glucose and lipid metabolism, thermogenesis, and the secretion of bioactive molecules that influence cardiac structure and function. Its proximity to the myocardium allows it to contribute directly to CVDs, including coronary artery disease, arrhythmias, and heart failure. In particular, excessive EAT has emerged as a significant factor in heart failure with preserved ejection fraction (HFpEF), the most common form of heart failure, especially in individuals with obesity and diabetes. Traditional metrics like body mass index (BMI) fail to capture the complexities of visceral fat, as patients with similar BMIs can exhibit varying CVD risks. EAT accumulation induces mechanical stress and fosters a pro-inflammatory and fibrotic environment, driving cardiac remodeling and dysfunction. Pharmacological modulation of EAT has shown promise in delivering cardiometabolic benefits. Recent advancements in diabetes therapies, such as SGLT2 inhibitors and GLP-1 receptor agonists, and antilipidemic drugs have demonstrated their potential in reducing pro-inflammatory cytokine production and improving glucose regulation, which directly influences EAT. These discoveries suggest that EAT could be a significant therapeutic target, though further investigation is necessary to elucidate its role in HFpEF and other CVDs. Recent advances have identified the prokineticin/PKR1 signaling pathway as pivotal in EAT development and remodeling. This pathway regulates epicardial progenitor cells (EPDCs), promoting angiogenesis while reducing EAT accumulation and metabolic stress on the heart, particularly under high-calorie conditions. Prokineticin, acting through its receptor PKR1, limits visceral adipose tissue growth, enhances insulin sensitivity, and offers cardioprotection by reducing oxidative stress and activating cellular survival pathways. In this review, we provide a comprehensive analysis of EAT's role in CVDs, explore novel therapeutic strategies targeting EAT, and highlight the potential of prokineticin signaling as a promising treatment for HFpEF, obesity, and diabetes.
Oleoylethanolamide-producing Lactobacillus paracasei F19 improves metabolic and behavioral disorders by restoring intestinal permeability and microbiota-gut-brain axis in high-fat diet-induced obese male mice Luisa Seguella, Chiara Corpetti, Jie Lu, Marcella Pesce, Silvia Basili Franzin, Irene Palenca, Aurora Zilli, Martina Vincenzi, Daniele Caprioli, Andreu Paytuví-Gallart, Walter Sanseverino, Sara Rurgo, Giovanni Sarnelli, Giuseppe Esposito Brain Behavior and Immunity, 2025 Metabolic and mood disorders elicited by chronic exposure of high-fat diet (HFD) are often associated with intestinal dysbiosis and persistent low-grade inflammation in the small intestine. This leads to remodeling of the epithelial barrier with disruption of the neuroepithelial circuits that control energy homeostasis by the gut-brain axis. Therefore, therapies that restoreintestinal microbial niche and barrier function are promising candidates to counter peripheral metabolic challenges that affect behaviors controlled by the brain. The endogenous oleoylethanolamine (OEA) was found to shape the intestinal microbiota profile towards a "lean-like phenotype", ameliorating pathological profiles of metabolic diseases. Further, OEA displays beneficial effects in several cognitive paradigms and preserves the epithelial barrier integrity, acting as an intestinal "gate-keeper". Here, we developed an "intestinal OEA factory" for the in-situ and controlled release of OEA by using a probiotic-based delivery system. We engineered the Lactobacillus paracasei F19 (LP) to express the human N-acylphosphatidylethanolamine-preferring phospholipase D (NAPEpld) gene and to produce OEA in response to dietary ultra-low oleate supply. We treated 12-week HFD male mice with oleate-probiotic formulations and assessed their impact on metabolic and behavioral dysfunctions, and microbiota-gut-brain signaling after 8 weeks of treatment. NAPE-expressing LP (pNAPE-LP) led to significant reduced weight loss and improved metabolic dysfunction in HFD-treated mice. Further, a parallel improvement in depressive- and anxiety-like phenotypes was associated with the duodenal barrier function retrieval, the restoration of the Firmicutes/Bacteroidetes ratio, and an increase in beneficial bacteria, such as Lactobacillus, Prevotella, and Parabacteroides. The HFD-driven changes both in the enteric and central nervous system were prevented by pNAPE-LP/oleate treatment. Collectively, our data suggest that these effects were mediated by the oleate-dependent release of OEA by pNAPE-LP since no significant effects were observed in HFD mice treated with the native probiotic alone (pLP). This oleate-regulated delivery system of OEA is a safe and efficient probiotic-based strategy for the treatment of metabolic syndrome and related behavioral disorders.
N-palmitoyl-d-glucosamine limits mucosal damage and VEGF-mediated angiogenesis by PPARα-dependent suppression of pAkt/mTOR/HIF1α pathway and increase in PEA levels in AOM/DSS colorectal carcinoma in mice Irene Palenca, Silvia Basili Franzin, Aurora Zilli, Luisa Seguella, Anna Troiani, Federico Pepi, Martina Vincenzi, Giuseppe Giugliano, Viviana Catapano, Italia Di Filippo, Giovanni Sarnelli, Giuseppe Esposito Phytotherapy Research, 2024 Chronic intestinal inflammation and neo‐angiogenesis are interconnected in colorectal carcinoma (CRC) pathogenesis. Molecules reducing inflammation and angiogenesis hold promise for CRC prevention and treatment. N‐Palmitoyl‐d‐glucosamine (PGA), a natural glycolipid analog with anti‐inflammatory properties, has shown efficacy against acute colitis. Micronized PGA (mPGA) formulations exhibit superior anti‐inflammatory activity. This study investigates the in vivo anti‐angiogenic and protective effects of mPGA in a mouse model of colitis‐associated CRC induced by azoxymethane/dextran sodium sulfate (AOM/DSS). CRC was induced in C57BL/6J mice using intraperitoneal azoxymethane followed by three cycles of 2.5% dextran sodium sulfate (DSS) in drinking water. Mice were treated with mPGA (30–150 mg/kg) with or without the PPARα inhibitor MK886 (10 mg/kg). At Day 70 post‐azoxymethane injection, mice underwent anesthetized endoscopic colon evaluation. Post‐mortem analysis of tumorigenesis and angiogenesis was performed using histological, immunohistochemical, and immunoblotting techniques. mPGA improved disease progression and survival rates in a dose‐ and PPARα‐dependent manner in AOM/DSS‐exposed mice. It reduced polyp formation, decreased pro‐angiogenic CD31, pro‐proliferative Ki67, and pro‐inflammatory TLR4 expression levels, and inhibited VEGF and MMP‐9 secretion by disrupting the pAkt/mTOR/HIF1α pathway. mPGA increased colon PEA levels, restoring anti‐tumoral PPARα and wtp53 protein expression. Given its lack of toxicity, mPGA shows potential as a nutritional intervention to counteract inflammation‐related angiogenesis in CRC.
MRAP2a Binds and Modulates Activity and Localisation of Prokineticin Receptor 1 in Zebrafish Maria Rosaria Fullone, Daniela Maftei, Martina Vincenzi, Roberta Lattanzi, Rossella Miele International Journal of Molecular Sciences, 2024 The prokineticin system plays a role in hypothalamic neurons in the control of energy homeostasis. Prokineticin receptors (PKR1 and PKR2), like other G-protein-coupled receptors (GPCRs) are involved in the regulation of energy intake and expenditure and are modulated by the accessory membrane protein 2 of the melanocortin receptor (MRAP2). The aim of this work is to characterise the interaction and regulation of the non-melanocortin receptor PKR1 by MRAP2a in zebrafish (zMRAP2a) in order to use zebrafish as a model for the development of drugs targeting accessory proteins that can alter the localisation and activity of GPCRs. To this end, we first showed that zebrafish PKR1 (zPKR1) is able to interact with both zMRAP2a and human MRAP2 (hMRAP2). This interaction occurs between the N-terminal region of zPKR1 and the C-terminal domain of zMRAP2a, which shows high sequence identity with hMRAP2 and a similar propensity for dimer formation. Moreover, we demonstrated that in Chinese hamster ovary (CHO) cells, zMRAP2a or hMRAP2 are able to modulate zPKR1 activation induced by zebrafish PK2 (zPK2) resulting in an impaired ERK and STAT3 activation.
Mapping the interaction site for β-arrestin-2 in the prokineticin 2 receptor R. Lattanzi, I. Casella, M.R. Fullone, M. Vincenzi, D. Maftei, R. Miele Cellular Signalling, 2024 G protein-coupled receptors (GPCRs) are a family of cell membrane receptors that couple and activate heterotrimeric G proteins and their associated intracellular signalling processes after ligand binding. Although the carboxyl terminal of the receptors is essential for this action, it can also serve as a docking site for regulatory proteins such as the β-arrestins. Prokineticin receptors (PKR1 and PKR2) are a new class of GPCRs that are able to activate different classes of G proteins and form complexes with β-arrestins after activation by the endogenous agonists PK2. The aim of this work was to define the molecular determinants within PKR2 that are required for β-arrestin-2 binding and to investigate the role of β-arrestin-2 in the signalling pathways induced by PKR2 activation. Our data show that PKR2 binds constitutively to β-arrestin-2 and that this process occurs through the core region of the receptor without being affected by the carboxy-terminal region. Indeed, a PKR2 mutant lacking the carboxy-terminal amino acids retains the ability to bind constitutively to β-arrestin-2, whereas a mutant lacking the third intracellular loop does not. Overall, our data suggest that the C-terminus of PKR2 is critical for the stability of the β-arrestin-2-receptor complex in the presence of PK2 ligand. This leads to the β-arrestin-2 conformational change required to initiate intracellular signalling that ultimately leads to ERK phosphorylation and activation.
Olfactory Neuron Substance P is Overexpressed in Parkinson's Disease Reflecting Gut Dysfunction Tommaso Schirinzi, Daniela Maftei, Piergiorgio Grillo, Roberta Bovenzi, Jacopo Bissacco, Clara Simonetta, Davide Mascioli, Henri Zenuni, Rocco Cerroni, Martina Vincenzi, Riccardo Maurizi, Francesco M. Passali, Stefano Di Girolamo, Massimo Ralli, Giuseppe Magliulo, Paola Tirassa, Alessandro Stefani, Nicola B. Mercuri, Roberta Lattanzi, Cinzia Severini Movement Disorders, 2023
Therapeutic Potential of Targeting Prokineticin Receptors in Diseases Martina Vincenzi, Amin Kremić, Appoline Jouve, Roberta Lattanzi, Rossella Miele, Mohamed Benharouga, Nadia Alfaidy, Stephanie Migrenne-Li, Anumantha G. Kanthasamy, Marimelia Porcionatto, Napoleone Ferrara, Igor V. Tetko, Laurent Désaubry, Canan G. Nebigil Pharmacological Reviews, 2023
Olfactory Neuron Prokineticin-2 as a Potential Target in Parkinson's Disease Tommaso Schirinzi, Daniela Maftei, Francesco M. Passali, Piergiorgio Grillo, Henri Zenuni, Davide Mascioli, Riccardo Maurizi, Laura Loccisano, Martina Vincenzi, Anna Maria Rinaldi, Massimo Ralli, Stefano Di Girolamo, Alessandro Stefani, Roberta Lattanzi, Cinzia Severini, Nicola B. Mercuri Annals of Neurology, 2023
Phytotherapics in COVID19: Why palmitoylethanolamide? Marcella Pesce, Luisa Seguella, Sara Cassarano, Laura Aurino, Walter Sanseverino, Jie Lu, Chiara Corpetti, Alessandro Del Re, Martina Vincenzi, Giovanni Sarnelli, Giuseppe Esposito Phytotherapy Research, 2021
