Research project 1
Cellular and Molecular Analysis of Hemato-Vascular Development
Principal Investigators:
Emanuele Azzoni, Silvia Brunelli
Study of the biology of embryonic/fetal hematopoietic stem cells and their precursors
- Genetic tracking of the contribution of embryonic and fetal hematopoietic precursors in transgenic murine models
- Imaging of hematopoietic precursors in their niche of emergence and prenatal maturation
- Ex vivo and in vivo functional studies to assess the lineage potential of fetal hematopoietic progenitors
- Single-cell transcriptomic studies to decipher the heterogeneity of embryonic and fetal hematopoietic stem cells and their precursors
Study of the role of the CXCL12-CXCR4-HMGB1 molecular axis in hemato-vascular development
- Identification of the functional role of the CXCL12-CXCR4 signaling axis in the generation and expansion of embryonic and fetal hematopoietic progenitors.
- Analysis of the impact of CXCL12-HMGB1-CXCR4 interaction as a mediator of pro-inflammatory signals in prenatal hematopoietic development.
- Functional studies on CXCR4 ligand variants in fetal tissue macrophage development.
- Definition of the cellular and molecular mechanisms through which modulation of the CXCR4 signal regulates angiogenesis (collaboration with UniMI)
Study of the role of stromal cells in the development and remodeling of blood vessels
- Use of organ-on-a-chip technology and next-generation microfluidic platforms to model the interaction between endothelial cells (EC) and mesenchymal stem cells (MSCs) or tissue-resident macrophages (TRMs)
- Real-time imaging of 3D vascular structures
- Analysis of biological parameters (vascular network complexity, perfusion, barrier function) and biomechanical parameters (tissue stiffness).
- Molecular analysis (single-cell transcriptomics) for the identification of key genes in EC-MSC and EC-TRM interaction
- Functional studies on the role of key genes in EC-MSC and EC-TRM interaction with a loss-of-function approach using siRNA and CRISPR/Cas9 techniques.
Publications
- Azzoni E*, Fantin A*, “Fetal liver hematopoiesis revisited: a precast hierarchy”, Nature Cardiovascular Research 2022 Oct:1:872-873. doi: 10.1038/s44161-022-00142-5.
- Tacconi C, Plein A, Colletto C, Villa E, Denti L, Barone C, Javanmardi Y, Moeendarbary E, Azzoni E, Fantin A, Ruhrberg C, “KIT is dispensable for physiological organ vascularisation in the embryo”, Angiogenesis 2022 Aug;25(3):343-353. doi: 10.1007/s10456-022-09837-6. Epub 2022 Apr 13.
- Barone C, Orsenigo R, Meneveri R, Brunelli S, Azzoni E* “One Size Does Not Fit All: Heterogeneity in Developmental Hematopoiesis”, Cells 2022 Mar 21;11(6):1061. doi: 10.3390/cells11061061
- Azzoni E*, Frontera V, Anselmi G, Rode C, James C, Deltcheva EM, Demian AS, Brown J, Barone C, Patelli A, Harman J, Nicholls M, Conway SJ, Morrissey E, Jacobsen SE, Sparrow DB, Harris AL, Enver T, de Bruijn MFTR*, “The onset of circulation triggers a metabolic switch required for endothelial to hematopoietic transition”, Cell Reports, 2021 Dec 14;37(11):110103. doi: 10.1016/j.celrep.2021.110103.
- Neo WH, Meng Y, Rodriguez-Meira A, Fadlullah M, Booth C, Azzoni E, Thongjuea S, de Bruijn MFTR, Jacobsen SE, Mead A, Lacaud G “Ezh2 is essential for the generation of functional yolk sac derived erythro-myeloid progenitors”, Nature Communications, 2021 Dec 2;12(1):7019. doi: 10.1038/s41467-021-27140-8.
Funding
- NextGeneration EU and Italian Ministry of University and Research (PRIN 2022 PNRR), “Novel roles for CXCR4 signaling in HEmatovascular development (CHEVron)”
- NextGeneration EU and Italian Ministry of University and Research (PRIN 2022), “Exploiting organ-on-a-chip technology to unravel the dynamic adaptation of microvasculature to biomechanical environmental cues (OC2AdMirE)”
Collaborations
- Alessandro Fantin, Prof. Matteo Chiara, Università degli Studi di Milano, Milano
- Rocco Piazza, Prof. Luca Mologni, Dipartimento di Medicina e Chirurgia, Università degli Studi di Milano-Bicocca
- Marco Bianchi, San Raffaele Scientific Institute, Milano
- Tullio Genova, Università degli Studi di Torino, Torino
- Georges Lacaud, University of Manchester, Manchester (United Kingdom)
- Charlotta Böiers, Lund University, Lund (Sweden)
- Sten Eirik Jacobsen, Karolinska Institutet, Stockholm (Sweden)
- Marella de Bruijn, University of Oxford (United Kindgom)
- Christiana Ruhrberg, UCL (University College London), London (United Kingdom)
- Ana Cumano, Institut Pasteur, Paris (France)
Research project 2
Modeling of Pediatric Myeloid Malignancies to Discover new Therapeutic Vulnerabilities
Principal Investigators:
Emanuele Azzoni
Study of the biology underlying Juvenile Myelomonocytic Leukemia (JMML) in transgenic and humanized murine models
- Generation and analysis of novel transgenic murine models of JMML to study its prenatal origins
- Definition of the cellular context of susceptibility to leukemic transformation by mutations responsible for JMML onset
- Single-cell multi-omics studies to define the molecular and epigenetic profile associated with JMML onset in embryonic and fetal hematopoietic progenitors
- Generation of a humanized murine model of JMML based on the xenotransplantation of bone marrow samples from pediatric patients into immunodeficient mice
Analysis of the functional role of the inflammatory response in JMML
- Characterization of the inflammatory response in transgenic and humanized murine models of JMML
- Analysis of the functional role of prenatal maternal inflammation in transgenic murine models of JMML
- Use of genetic and pharmacological approaches to study the role of genes involved in molecular pathways of cellular stress response and pro-inflammatory response in transgenic murine models of JMML
- Use of genetic and pharmacological approaches to study the role of genes involved in molecular pathways of cellular stress response and pro-inflammatory response in humanized murine models of JMML
Generation and analysis of murine models of pediatric leukemias with KMT2A (MLL) rearrangements
- Use of CRISPR/Cas9 technology in transgenic murine models to engineer Mll-involved translocations in embryonic and fetal hematopoietic stem and progenitor cells
- Analysis of the leukemic phenotype in relation to differences in hematopoietic stem/progenitor cells targeted by Mll rearrangements involved in pediatric Acute Myeloid Leukemia (AML) and Acute Lymphoblastic Leukemia (ALL)
- Study of the molecular profile (transcriptomics, proteomics) of leukemic cells in relation to the developmental state of hematopoietic stem/progenitor cells acquiring Mll rearrangements
Publications
- Crespiatico I, Zaghi M, Mastini C, D’Aliberti D, Mauri M, Mercado CM, Fontana D, Spinelli S, Crippa V, Inzoli E, Manghisi B, Civettini I, Ramazzotti D, Sangiorgio V, Gengotti M, Brambilla V, Aroldi A, Banfi F, Barone C, Orsenigo R, Riera L, Riminucci M, Corsi A, Breccia M, Morotti A, Cilloni D, Roccaro AM, Sacco A, Stagno F, Serafini M, Mottadelli F, Cazzaniga G, Pagni F, Chiarle R, Azzoni E, Sessa A, Gambacorti-Passerini C, Elli EM, Mologni L, Piazza R, “First-hit SETBP1 mutations cause a myeloproliferative disorder with bone marrow fibrosis”, Blood 2024 Apr 4;143(14):1399-1413. doi: 10.1182/blood.2023021349.
- Barone C, Orsenigo R, Cazzola A, D’Errico E, Patelli A, Quattrini G, Vergani B, Bombelli S, De Marco S, D’Orlando C, Bianchi C, Leone BE, Meneveri R, Biondi A, Cazzaniga G, Rabbitts TH, Brunelli S and Azzoni E*. Hematopoietic Stem Cell (HSC)-Independent Progenitors Are Susceptible to Mll-Af9-Induced Leukemic Transformation. Cancers. 2023 Jul 14;15(14):3624. https://doi.org/10.3390/cancers15143624
- Zaghi M, Banfi F, Massimino L, Volpin M, Bellini E, Brusco S, Merelli I, Barone C, Bruni M, Bossini L, Lamparelli L, Pintado L, D’Aliberti S, Spinelli S, Mologni L, Colasante G, Ungaro F, Cioni JM, Azzoni E, Piazza R, Montini E, Broccoli V, Sessa A, “Balanced SET levels favor the correct enhancer repertoire during cell fate acquisition”, Nature Communications, 2023 Jun 3;14(1):3212. doi: 10.1038/s41467-023-39043-x.
- Cazzola A, Cazzaniga G, Biondi A, Meneveri R, Brunelli S, Azzoni E*, “Prenatal origin of pediatric leukemia: lessons from hematopoietic development”, Frontiers in Cell and Developmental Biology, 2021 doi: https://doi.org/10.3389/fcell.2020.618164. *: corresponding author
Funding
- WorldWide Cancer Research, “Targeting inflammatory pathways in juvenile myelomonocytic leukemia”
- CARIPLO-Telethon Alliance, “Novel targets modulating the inflammatory response in juvenile myelomonocytic leukemia”
Collaborations
- Giuseppe Gaipa, Fondazione Tettamanti, Monza
- Rocco Piazza, Prof. Luca Mologni, Dipartimento di Medicina e Chirurgia, Università degli Studi di Milano-Bicocca
- Charlotta Böiers, Lund University, Lund (Sweden)
- Antonella Ronchi, BtBs, Università degli Studi di Milano-Bicocca, Milano
- Laura Belver, Josep Carreras Leukemia Research Institute, Barcelona (Spain)
- Ciprian Tomuleasa, Iuliu Hatieganu University of Medicine and Pharmacy, Cluj Napoca (Romania)
- Cheng-Kui Qu, Emory University School of Medicine, Atlanta (USA)
Research project 3
Deciphering Immune Modulation in Ectopic Ossification: Insights from Fibrodysplasia Ossificans Progressiva (FOP)
Principal Investigators:
Silvia Brunelli
Develop Advanced Experimental Models
- Establish and optimize a mouse preclinical model of FOP (Acvr1R206Hlox/lox;Gt(ROSA26)SorCreERT2) to study immune modulation and tissue remodeling.
- Develop innovative tools and technologies, including 3D artificial niches, to mimic the microenvironment of ectopic ossification sites and study cellular responses.
Characterize Immune Modulation in FOP
- Utilize a multidisciplinary approach to dissect the role of immune cells, particularly macrophages, in the pathogenesis of FOP.
- Investigate the heterogeneity of immune responses in the microenvironment of ectopic ossification sites.
- Determine the impact of genetic mutations and environmental triggers on immune cell dynamics in FOP.
Elucidate Osteogenic Progenitor Modulation by Macrophages
- Explore the crosstalk between macrophages and osteogenic progenitors during the initiation and progression of ectopic ossification.
- Determine the mechanisms by which immune modulation influences the fate and function of osteogenic progenitor cells.
- Investigate the role of cytokines and signaling pathways in mediating macrophage-osteogenic progenitor interactions.
Publications
- Cappato, S., Gamberale, R., Bocciardi, R., Brunelli, S., 2020. Genetic and Acquired Heterotopic Ossification: A Translational Tale of Mice and Men. Biomedicines 8.
- Convente, M.R., Chakkalakal, S.A., Yang, E., Caron, R.J., Zhang, D., Kambayashi, T., Kaplan, F.S., Shore, E.M., 2017. Depletion of Mast Cells and Macrophages Impairs Heterotopic Ossification in an Acvr1R206H Mouse Model of Fibrodysplasia Ossificans Progressiva. J Bone Miner Res 33, 269-282.
- Kaplan, F.S., Xu, M., Seemann, P., Connor, J.M., Glaser, D.L., Carroll, L., Delai, P., Fastnacht-Urban, E., Forman, S.J., Gillessen-Kaesbach, G., Hoover-Fong, J., Koster, B., Pauli, R.M., Reardon, W., Zaidi, S.A., Zasloff, M., Morhart, R., Mundlos, S., Groppe, J., Shore, E.M., 2009. Classic and atypical fibrodysplasia ossificans progressiva (FOP) phenotypes are caused by mutations in the bone morphogenetic protein (BMP) type I receptor ACVR1. Hum Mutat 30, 379-390.
- Pignolo, R.J., Bedford-Gay, C., Liljesthrom, M., Durbin-Johnson, B.P., Shore, E.M., Rocke, D.M., Kaplan, F.S., 2016. The Natural History of Flare-Ups in Fibrodysplasia Ossificans Progressiva (FOP): A Comprehensive Global Assessment. J Bone Miner Res 31, 650-656.
- Théret, M., Mounier, R., Rossi, F., 2019. The origins and non-canonical functions of macrophages in development and regeneration. Development 146, dev156000-156014.
- Tirone, M., Giovenzana, A., Vallone, A., Zordan, P., Sormani, M., Nicolosi, P.A., Meneveri, R., Gigliotti, C.R., Spinelli, A.E., Bocciardi, R., Ravazzolo, R., Cifola, I., Brunelli, S., 2019. Severe Heterotopic Ossification in the Skeletal Muscle and Endothelial Cells Recruitment to Chondrogenesis Are Enhanced by Monocyte/Macrophage Depletion. Front Immunol 10, 1640.
- Wang, H., Shore, E.M., Pignolo, R.J., Kaplan, F.S., 2018. Activin A amplifies dysregulated BMP signaling and induces chondro-osseous differentiation of primary connective tissue progenitor cells in patients with fibrodysplasia ossificans progressiva (FOP). Bone 109, 218-224.
Funding
- Marie Skodowska-Curie Actions (MSCA) Innovative Training Networks (ITN) H2020-MSCA-ITN-2019: REcreating the ideal Niche: environmental control Of cell Identity in Regenerating and diseased muscles. RENOIR (grant agreement No 860034)
- Associazione FOP Italia . “Caratterizzazione cellulare e molecolare dei macrofagi polarizzati infiltranti durante l’insorgenza dell’ossificazione eterotopica in un modello murino di Fibrodisplasia Ossificans Progressiva”
- Italian Ministry of University-Fondo per il Programma Nazionale di Ricerca e Progetti di Rilevante Interesse Nazionale (PRIN) – Environmental control oF Ectopic oSTeOgenesis in Fibrodysplasia Ossificans Progressiva: from mouse to chip and back-EFESTO (2022TR9N4R)
- Fondazione Telethon – Exploring the Role of SPP1 in Macrophage-Osteogenic Cell Crosstalk and its involvement in Heterotopic Ossification in Fibrodysplasia Ossificans Progressiva (GMR24T1043)
Collaborations
- Renata Bocciardi, University of Genoa, Genoa, Italy
- Benedicte Chazaud, Université Claude Bernard Lyon 1, Lyone, France
- Gonzalo Sánchez-Duffhues, Health Research Institute of Asturias, Oviedo, Spain
- Patrizia Rovere Querini, San Raffaele Scientific Institute and San Raffaele University, Milano, Italy
- Angelo A. Manfredi, San Raffaele Scientific Institute and San Raffaele University, Milano, Italy
- Marco Bianchi, San Raffaele Scientific Institute and San Raffaele University, Milano, Italy
- Gabriella Minchiotti, Institute of Genetics and Biophysics “Adriano Buzzati Traverso”, CNR, Naples, Italy
Latest Publications
We are pleased to share our work published in peer-reviewed international journals indexed in Pubmed and Scopus
Passion for blood
From developmental hematopoiesis to the remodeling vascular niche.
We are a multi-disciplinary laboratory that is committed to pursue high quality scientific research in the field of hematopoietic and vascular cell biology