Deep learning for predicting stem cell efficiency for use in beta cell differentiation Franziska J. Schöb, Alexander Binder, Valentina Zamarian, Valeria Sordi, Hanne Scholz, Anders Malthe-Sørenssen, Dag Kristian Dysthe Scientific Reports, 2026 Recent clinical trial data show curative potential of cell therapy for diabetes, however the cells required are a bottleneck. Cell differentiation exhibits substantial variability, even among clones of stem cells generated from the same patient. Human experts struggle to see the difference between highly- and lowly-efficient cell clones early. We therefore propose an image-based deep learning model to guide the selection of the most efficient clones. We apply different deep learning models to learn the morphological differences between good and bad stem cell clones and classify them based on phase-contrast imaging. To gain insight into the learned features, we use layer-wise relevance propagation, and Fourier-based frequency analysis. Using an EfficientNet-V2-S model, we obtain a novel early prediction for the outcome of the differentiation process from patient-derived stem cells to [Formula: see text] -cells using imaging. Clone level accuracy is 96.7 % at 53 hours after start of differentiation. The analysis of learned features shows that the structure of the cell population is an important predictive feature. This study is a proof-of-concept that deep learning combined with label-free imaging can be highly predictive and guide selection of stem cell clones, thereby reducing cost of [Formula: see text] -cell production.
Gene Expression at the Pluripotency Stage Predicts Pancreatic Endocrine Differentiation in iPSC Clones Valentina Zamarian, Laura Monaco, Manuela Marras, Chiara Ceriani, Silvia Pellegrini, Lorenzo Piemonti, Valeria Sordi Stem Cell Reviews and Reports, 2026 Induced pluripotent stem cell (iPSC)-derived β-like cells hold great promise for cell replacement therapy in type 1 diabetes. However, the reprogramming process generates iPSC clones with variable differentiation capacity, hindering the selection of optimal cell lines. This study aimed to identify an early-stage transcriptional signature capable of predicting the β cell differentiation potential of donor-matched iPSC clones. Eleven iPSC clones derived from a single donor were differentiated to the definitive endoderm (DE) stage; six were further driven toward pancreatic progenitors (PP) and insulin-producing cells. Differentiation efficiency was evaluated by flow cytometry and qPCR at iPSC, DE, PP, and β cell stages. At the pluripotent stage, expression profiling of 770 genes related to pluripotency and trilineage specification was performed to identify predictive molecular markers. Transcriptomic analysis segregated the clones into two groups (Gr1 and Gr2) with significantly different differentiation outcomes. Gr2 clones exhibited superior DE efficiency (Cxcr4⁺: 90.1 ± 5.6% vs. 79.8 ± 3.6%; P = 0.027) and higher expression of PP markers (Pdx1⁺, Nkx6.1⁺, and double-positive cells; P ≤ 0.05). At the β cell stage, Gr2 clones showed increased frequencies of Pdx1⁺/Ins⁺ and Nkx6.1⁺/Ins⁺ cells (P ≤ 0.05), along with enhanced glucose-stimulated insulin secretion. A set of 73 differentially expressed genes, enriched in pathways related to naïve/primed pluripotency, endoderm commitment, and metabolism, was identified. From this, a ten-gene signature validated by qPCR strongly correlated with pancreatic marker expression at all stages. An early gene expression signature at the pluripotent stage predicts the pancreatic endocrine differentiation potential of iPSC clones. This molecular screening approach may enable rapid preselection of high-performing clones, thereby accelerating the development of personalized stem cell–based therapies for diabetes. Cellular reprogramming is a fundamental tool in regenerative medicine but often produces iPSC clones with heterogeneous differentiation potential. Identifying the most suitable clones typically requires time-consuming assays and prolonged in vitro testing. This study presents a streamlined transcriptomic approach to predict, at the pluripotent stage, the differentiation efficiency of iPSC clones into pancreatic endoderm and insulin-producing cells, enabling early selection of high-performing lines for the development of diabetes cell therapy.
Vav1 Sustains the Expression of Insulin, PDX1 and miR-375 During Differentiation of hiPSCs to β Cells: A Potential Target to Improve the In Vitro Generation of Insulin-Producing Cells Marina Pierantoni, Valentina Zamarian, Federica Brugnoli, Silvia Grassilli, Laura Monaco, Marcello Dell’Aira, Valeria Sordi, Valeria Bertagnolo Tissue Engineering and Regenerative Medicine, 2026 Backround: Human-induced pluripotent stem cells (hiPSCs) have emerged as a promising source of transplantable insulinproducing cells (IPCs) to restore insulin levels in Type 1 Diabetes (T1D) patients. Despite progress, obtaining fully functional β cells from hiPSCs remains challenging, underscoring the need to better understand the intracellular mechanisms involved. We investigated here the potential role of Vav1, a multidomain protein that we identified as crucial for the maturation of human biliary stem cells (hBTSCs) into β-like cells and in the trans-differentiation of pancreatic adenocarcinoma (PDAC) cells into IPCs; Methods: Levels and subcellular localization of Vav1 were investigated throughout a seven-step differentiation process of hiPSCs to β cells. Vav1expression was forcedly modulated in pancreatic progenitors, and the potential effects were evaluated on insulin production and on PDX1, miR-375, and Akt, key regulators of β cells generation; RESULTS. Vav1 showed dynamic modulation, with pancreatic precursor cells requiring adequate levels of the protein to generate IPCs. Results: Vav1 sustains the expression of PDX1, a primary regulator of insulin expression, and of its target miR-375, essential for determining β cell mass. Furthermore, Vav1 reduction correlated with increased activation of Akt, which regulates cell survival and insulin secretion in β cells and is down-regulated by miR- 375. Conclusion: Our findings suggest the existence of a Vav1/PDX1/miR-375/Akt axis as part of the complex network orchestrating the generation of functional β cells. These insights indicate that strategies aimed at specifically modulating Vav1 levels may positively impact the generation of IPCs in vitro and, ultimately, β cell replacement therapy for T1D.
Induction of immune education in type 1 diabetes through controlled allogeneic islet rejection at onset: a monocentric open-label pilot study Lorenzo Piemonti, Andrea Mario Bolla, Amelia Caretto, Raffaella Melzi, Alessia Mercalli, Valeria Sordi, Paolo Monti, Paola Magistretti, Vito Lampasona, Ilaria Marzinotto, Paola Maffi, Miriam Ramondetta, Nicoletta Cagni, Erica Pedone, Davide Catarinella, Massimo Cardillo, Rossana Caldara, Emanuele Bosi Eclinicalmedicine, 2025 Background: Current immunotherapies for type 1 diabetes (T1D) have shown limited success in durably preserving β-cell function. We tested a novel strategy that repurposes allogeneic islet transplantation not for metabolic replacement, but as a platform for antigen-specific immune education. Methods: In this monocentric, open-label pilot study (April 2015-April 2023), six patients with recent-onset T1D received a minimal islet mass (median 3452 IEQ/kg; range 2980-4050) combined with short-term immunomodulation (ATG, transient mTOR inhibition, and G-CSF). The transplanted islet mass was intentionally insufficient for metabolic replacement. The primary endpoint was the change in stimulated 2-h C-peptide AUC at 52 weeks. Exploratory endpoints included immune cell phenotyping, cytokine/chemokine profiling, miR-375 release kinetics, analysis of islet-related autoantibodies, and monitoring of donor-specific HLA antibodies. ClinicalTrials.gov Identifier: NCT02505893. Findings: T cells specific for GAD65 was detected. Interpretation: This study introduces a paradigm shift in the use of islet transplantation-transforming it from a metabolic intervention into a tolerogenic stimulus through controlled antigen exposure. Further potentiation with stem-cell-derived islets may enable improved matching, graft modification, and iterative antigen delivery. Funding: Supported by Fondazione Italiana Diabete (FID). The funder had no role in study design, data collection, data analysis, interpretation, manuscript preparation, or decision to publish.
Liraglutide Treatment Reverses Unconventional Cellular Defects in Induced Pluripotent Stem Cell–Derived β-Cells Harboring a Partially Functional WFS1 Variant Silvia Torchio, Gabriel Siracusano, Federica Cuozzo, Valentina Zamarian, Silvia Pellegrini, Fabio Manenti, Riccardo Bonfanti, Giulio Frontino, Valeria Sordi, Raniero Chimienti, Lorenzo Piemonti Diabetes, 2025 Wolfram syndrome 1 (WS1) is a rare genetic disorder caused by WFS1 variants that disrupt wolframin, an endoplasmic reticulum-associated protein essential for cellular stress responses, Ca2+ homeostasis, and autophagy. Here, we investigated how the c.316-1G>A and c.757A>T WFS1 mutations, which yield partially functional wolframin, affect the molecular functions of β-cells and explored the therapeutic potential of the glucagon-like peptide 1 receptor (GLP-1R) agonist liraglutide. Pancreatic β-cells obtained from patient-derived induced pluripotent stem cells (iPSCs) carrying this WFS1 variant exhibited reduced insulin processing and impaired secretory granule maturation, as evidenced by proinsulin accumulation and decreased prohormone convertase PC1/3. Moreover, they exhibited dysregulated Ca2+ fluxes due to altered transcription of Ca2+-related genes, including CACNA1D, and significantly reduced SNAP25 levels, leading to uncoordinated oscillations and poor glucose responsiveness. Affected cells also showed increased autophagic flux and heightened susceptibility to inflammatory cytokine-induced apoptosis. Notably, liraglutide treatment rescued these defects by normalizing Ca2+ handling, enhancing insulin processing and secretion, and reducing apoptosis, likely through modulation of the unfolded protein response. These findings underscore the importance of defining mutation-specific dysfunctions in WS1 and support targeting the GLP-1/GLP-1R axis as a therapeutic strategy. Article Highlights The molecular basis of WFS1-related mutations remains poorly investigated, and no definitive therapies exist for Wolfram syndrome 1. We dissected the molecular defects associated with c.316-1G>A and c.757A>T WFS1 mutations in patient-derived induced pluripotent stem cell islets and analyzed whether they are potential therapeutic targets of the glucagon-like peptide 1 receptor agonist liraglutide. We found impaired insulin granule maturation, altered Ca2+ fluxes, increased autophagic activity, and heightened susceptibility to inflammatory apoptosis in mutated cells. Liraglutide restored critical β-cell functions suggesting a route for personalized therapy based on WFS1 mutations.
SID/SIEDP expert consensus on optimizing clinical strategies for early detection and management of wolfram syndrome Giulio Frontino, Maurizio Delvecchio, Sabrina Prudente, Valeria Daniela Sordi, Piero Barboni, Alessandra Di Giamberardino, Alessandra Rutigliano, Silvia Pellegrini, Amelia Caretto, Maria Lucia Cascavilla, Riccardo Bonfanti, Giuseppe D’Annunzio, Fortunato Lombardo, Lorenzo Piemonti Journal of Endocrinological Investigation, 2025 Wolfram Syndrome (WFS) is a rare, multisystemic, degenerative disease leading to premature death. Clinical and genetic heterogeneity makes WFS diagnosis and management challenging. The Italian Society of Diabetes (SID) and the Italian Society for Pediatric Endocrinology and Diabetology (SIEDP) convened an expert panel of professional healthcare practitioners to provide up-to-date knowledge about the pathophysiology, clinical presentation and treatment of WFS, and recommendations for the earlydetection and optimal disease management. The consensus recommends the revision of diagnostic protocols to include genetic testing and comprehensive multidisciplinary evaluations to ensure accurate diagnosis of WFS, advocates for personalized management plans tailored to the unique needs of each patient, with an emphasis on exploring new potential drug therapies. A holistic care model that addresses the medical, psychological, and social challenges faced by patients with WFS and their families is strongly endorsed. The opinion underscores the importance of educating healthcare professionals about WFS to enhance early diagnosis and intervention, aiming to improve outcomes for patients through practical and evidence-based clinical strategies.
Blockade of CD155 and CD276 by Monoclonal Antibodies Fosters Immune Tolerance and Promotes Stable Engraftment of iPSC-Derived Islets in Allogeneic Humanized Mice G. Siracusano, F. Deambrogio, V. Sordi, M. Malnati, L. Piemonti, R. Chimienti Transplant International, 2025 Induced pluripotent stem cell (iPSC)-derived pancreatic islets represent a promising therapeutic approach for restoring insulin independence in type 1 diabetes (T1D). However, their clinical success remains critically dependent on overcoming rejection mediated by innate and adaptive immune responses. Current immunosuppressive therapies pose significant long-term risks and only partially control alloimmune and autoimmune reactions. Targeted immunomodulation using monoclonal antibodies is a safer, more precise alternative. Here, we explored the impacts of blocking CD276 (B7-H3) and CD155 (PVR), activating ligands involved in immune recognition and regulation, on the survival and in vivo maturation of iPSC-derived endocrine progenitors (EPs) into functional pancreatic islets. Using a humanized mouse model, we demonstrated that dual blockade of CD276 and CD155 markedly reduced NK cell-mediated graft rejection, prevented CD14 + monocyte activation, and limited overall immune infiltration. In addition, CD155 blockade increased PD-1 levels on activated CD8 + T cells and significantly enhanced regulatory T cell (Treg) expansion and function, thereby promoting graft tolerance. Combined treatment prolonged engraftment and facilitated the maturation of EPs into functional, insulin-secreting cells, as indicated by increased human C-peptide levels and glucose responsiveness 4 weeks post-transplantation. Our findings highlight CD276/CD155 blockade as a novel immunomodulatory strategy to support tolerance and the functional maturation of iPSC-derived pancreatic grafts in T1D.
Implications of noncoding regulatory functions in the development of insulinomas Mireia Ramos-Rodríguez, Marc Subirana-Granés, Richard Norris, Valeria Sordi, Ángel Fernández, Georgina Fuentes-Páez, Beatriz Pérez-González, Clara Berenguer Balaguer, Helena Raurell-Vila, Murad Chowdhury, Raquel Corripio, Stefano Partelli, Núria López-Bigas, Silvia Pellegrini, Eduard Montanya, Montserrat Nacher, Massimo Falconi, Ryan Layer, Meritxell Rovira, Abel González-Pérez, Lorenzo Piemonti, Lorenzo Pasquali Cell Genomics, 2024
Faecal microbiota transplantation halts progression of human new-onset type 1 diabetes in a randomised controlled trial Pieter de Groot, Tanja Nikolic, Silvia Pellegrini, Valeria Sordi, Sultan Imangaliyev, Elena Rampanelli, Nordin Hanssen, Ilias Attaye, Guido Bakker, Gaby Duinkerken, Antoinette Joosten, Andrei Prodan, Evgeni Levin, Han Levels, Bartjan Potter van Loon, Arianne van Bon, Catherina Brouwer, Sytze van Dam, Suat Simsek, Daniel van Raalte, Frank Stam, Victor Gerdes, Roel Hoogma, Martin Diekman, Martin Gerding, Cees Rustemeijer, Bernadette de Bakker, Joost Hoekstra, Aeilko Zwinderman, Jacques Bergman, Frits Holleman, Lorenzo Piemonti, Willem De Vos, Bart Roep, Max Nieuwdorp Gut, 2021
Islet allotransplantation in the bone marrow of patients with type 1 diabetes: A pilot randomized trial Paola Maffi, Rita Nano, Paolo Monti, Raffaella Melzi, Valeria Sordi, Alessia Mercalli, Silvia Pellegrini, Maurilio Ponzoni, Jacopo Peccatori, Carlo Messina, Angela Nocco, Massimo Cardillo, Marina Scavini, Paola Magistretti, Claudio Doglioni, Fabio Ciceri, Stef J. Bloem, Bart O. Roep, Antonio Secchi, Lorenzo Piemonti Transplantation, 2019
Study of 2009 H1N1 pandemic influenza virus as a possible causative agent of diabetes I. Capua, A. Mercalli, Aurora Romero-Tejeda, M. Pizzuto, S. Kasloff, V. Sordi, I. Marzinotto, V. Lampasona, E. Vicenzi, C. De Battisti, R. Bonfanti, A. Rigamonti, C. Terregino, C. Doglioni, G. Cattoli, L. Piemonti Journal of Clinical Endocrinology and Metabolism, 2018
Stem cells to restore insulin production and cure diabetes V. Sordi, S. Pellegrini, M. Krampera, P. Marchetti, A. Pessina, G. Ciardelli, G. Fadini, C. Pintus, G. Pantè, L. Piemonti Nutrition Metabolism and Cardiovascular Diseases, 2017
Duodenal mucosa of patients with type 1 diabetes shows distinctive inflammatory profile and microbiota Silvia Pellegrini, Valeria Sordi, Andrea Mario Bolla, Diego Saita, Roberto Ferrarese, Filippo Canducci, Massimo Clementi, Francesca Invernizzi, Alberto Mariani, Riccardo Bonfanti, Graziano Barera, Pier Alberto Testoni, Claudio Doglioni, Emanuele Bosi, Lorenzo Piemonti Journal of Clinical Endocrinology and Metabolism, 2017
The human pancreas as a source of protolerogenic extracellular matrix scaffold for a new-generation bioartificial endocrine pancreas Andrea Peloso, Luca Urbani, Paolo Cravedi, Ravi Katari, Panagiotis Maghsoudlou, Mario Enrique Alvarez Fallas, Valeria Sordi, Antonio Citro, Carolina Purroy, Guoguang Niu, John P. McQuilling, Sivanandane Sittadjody, Alan C. Farney, Samy S. Iskandar, Joao P. Zambon, Jeffrey Rogers, Robert J. Stratta, Emmanuel C. Opara, Lorenzo Piemonti, Cristina M. Furdui, Shay Soker, Paolo De Coppi, Giuseppe Orlando Annals of Surgery, 2016
β-cell transplantation in diabetes mellitus Diabetes Research Institue, Ospidale San Raffaele, Milano; Universita degli Studi dell'Insubria, Varese, S Pellegrini, V Sordi, L Piemonti Diabetes Mellitus, 2013
Role of CCL2/MCP-1 in islet transplantation Raffaella Melzi, Alessia Mercalli, Valeria Sordi, Elisa Cantarelli, Rita Nano, Paola Maffi, Giovanni Sitia, Luca G. Guidotti, Antonio Secchi, Ezio Bonifacio, Lorenzo Piemonti Cell Transplantation, 2010
Bone marrow as an alternative site for islet transplantation Elisa Cantarelli, Raffaella Melzi, Alessia Mercalli, Valeria Sordi, Giuliana Ferrari, Carsten Werner Lederer, Emanuela Mrak, Alessandro Rubinacci, Maurilio Ponzoni, Giovanni Sitia, Luca G. Guidotti, Ezio Bonifacio, Lorenzo Piemonti Blood, 2009
