OASI: the Asplenia Observatory for Integrated Studies

A Multidisciplinary Challenge toward an Integrated Approach to Asplenia

This collaborative project relies on partnerships among three distinct yet interconnected fields:

• Clinics,
• biological sciences,
• computer science.

Background

Asplenia arises from either the surgical removal or intrinsic dysfunction of the spleen and is linked to a wide spectrum of hematological, oncological, immunological, and congenital disorders. Patients with asplenia face severe and potentially life-threatening complications, including heightened susceptibility to infections due to immune dysfunction and an increased risk of thrombotic events caused by coagulative and vascular imbalances. These complications profoundly affect long-term health outcomes, leading to greater disability and significant economic burdens on public health systems.

Despite its impact, research on asplenia lacks a unified, systematic approach, largely due to the multiplicity of diseases underlying this condition and to inter-individual variability. Existing studies are often confined to small, disease-specific cohorts, limiting the development of broad, evidence-based clinical guidelines.

The OASI Research Biobank

Asplenia represents a multidisciplinary challenge that requires an exhaustive approach to address its complexities effectively. To address these challenges, the OASI Biobank initiative was conceived to provide comprehensive resources for biomedical research on asplenia by collecting, preserving, and analyzing biological samples and related data from asplenic patients.

The OASI Biobank holds as defining features the transversal approach and the availability of clinical records updated over time. As asplenia occurs in many different clinical contexts, unlike traditional disease-oriented biobanks, the OASI Biobank addresses multiple pathologies intersecting this clinical condition. This innovative perspective enables research into the complex relationships between asplenia and its associated disorders. By broadening our understanding of these intricate interactions, the OASI Biobank aims to improve clinical outcomes and advance personalized medical interventions for asplenic patients.

The OASI Biobank strives to operate on a national scale as a centralized infrastructure through the close collaboration of the Italian Network for Asplenia (INA), coordinated by the “Luigi Vanvitelli” University Hospital in Naples. INA is the first and unique clinical network in Italy and Europe dedicated exclusively to the care of asplenic patients. Central to INA’s efforts was establishing an Observational Clinical Database, which compiles high-quality, prospective, patient-level data.

Through an access-controlled interface with the INA Database, the OASI Biobank incorporates comprehensive clinical data of donors. This seamless integration provides a continuously updated and robust resource that enables the cross-referencing and triangulation of metadata and analytical information from biobank donors with detailed clinical records.

The OASI Research Biobank seeks to facilitate multidisciplinary, multicenter, large-scale integrated studies aimed to fill critical gaps in asplenia biomedical research. Furthermore, OASI will promote bioinformatics innovation by leveraging computational biology to provide insights into asplenia.

Research linked to the OASI Biobank: the ABCare project.

The ABCare project relies on:

1) examination of biological material of donors to generate analytical data (at cellular, sub-cellular, and molecular levels using lab-scale and high-throughput approaches).

2) computational analysis (data integration, statistical testing of longitudinal data, correlation analysis, and comparative association) on:

• analytical data (including omics)
• metadata
• biometric, clinical, and laboratory information (retrospective and prospective data with quantitative and/or qualitative variables)

to perform:

observational and experimental studies intersecting multiple diseases:

• cross-sectional research (multiple experimental variables at a given instance analyzed in conjunction with retrospective clinical data)
• longitudinal survey (few experimental variables collected over time analyzed with retrospective and prospective clinical information). Analysis

with the aim of:

identifying clinical biomarkers of disease severity and prognostic tools for splenectomy evaluation and asplenia management:

• Risk biomarkers – for the assessment of long-term thrombotic and infectious risk in asplenic patients.
• Prognostic biomarkers – to predict the course of asplenia, to assess its severity and progression, regardless of the underlying disease.
• Screening biomarkers – for the subclinical assessment of splenic function.
• Predictive biomarkers – to develop risk scores to evaluate and prevent criticism of the recognized consequences and complications associated with therapeutic splenectomy.

Funding:

“ABCare: The Asplenia Biobanking Community: from Analytes to theRapEutic decision making”.

MUR PRIN 2022. Macrosettore LS-Life Sciences, Settore ERC LS7 “Prevention, Diagnosis and Treatment of Human Diseases”, Codice progetto 2022Y59MHL_LS7_PRIN2022.

Biobank Governance:

Biobank Coordinator: Dott.ssa Laura Casalino
Biobank Investigator: Dott.ssa Marcella Vacca

External Partners and People:

“Luigi Vanvitelli” University Hospital of Naples:

Prof.ssa Maddalena Casale

Institute for High-Performance Computing and Networking, ICAR-CNR.

Dott.ssa Ilaria Granata

Dott. Maurizio Giordano

Profiling the Mecp2 allelic expression in mouse brain development as a premise for its therapeutic reactivation in RTT.

The X-linked MECP2 gene is extensively studied in bridging epigenetics with human health. MECP2-associated diseases suggest that mammalian cells are susceptible to MeCP2 dosage. In females, heterozygous mutations cause Rett syndrome (RTT, OMIM: 312750), a severe neurodevelopmental disorder, while in males, hemizygous mutations lead to a more severe neonatal encephalopathy. Conversely, proximal Xq28 duplication syndrome (OMIM: 300260 300815), a functional Xq28 disomy involving the MECP2 gene, mainly affects males and results in neurodegeneration. These clinical findings suggest that MeCP2 functions are crucial in the intersection between neurodevelopmental and neurodegenerative disorders.

Moreover, compared to males, females better tolerate the adversities of X chromosome-related genetic diseases due to the neuroprotective action of the Lyon effect. This epigenetic process ensures dosage compensation between genders through X chromosome inactivation (XCI). XCI involves the random silencing of one of the X chromosomes during early embryonic development, with stable epigenetic mechanisms clonally inherited. Due to XCI, Mecp2 prevalently shows monoallelic expression: patients are an epigenetic mosaic of cells expressing either the mutant or the healthy allele, whose proportion in pathogenic tissues correlates with disease severity. However, evidence suggests that the murine Mecp2 may behave as a facultative lineage-specific escape from XCI that can be reactivated through XCI counteracting compounds. Therefore, we established an easy-to-use approach to monitor the spatial and temporal inactivation/activation status of Mecp2 and distinguish the allelic contribution to the MecP2 protein expression at a single-cell level. We developed female double knock-in mice with a two-color fluorescent reporter system tagging each of the Mecp2 alleles. Among major advantages, this transgenic strain preserves the physiological XCI, thus allowing the tracing of the allelic contribution and the parent-of-origin effect in the expression of MeCP2 protein. Our preliminary evidence suggests that Mecp2 epigenetic mosaicism in the brain might hide elusive details, namely that Mecp2 shows a spatially and developmentally regulated biallelic expression in a restricted portion of brain cells and areas. In perspective, such findings could foster reconsideration of some unsolved issues related to the MECP2-dependent Rett syndrome, while providing new paths to epigenetic manipulations for therapeutic purposes, aimed at the specific reactivation of the MeCP2-locus.

1. Casalino L, Talotta F, Matino I, Verde P. FRA-1 as a Regulator of EMT and Metastasis in Breast Cancer. Int J Mol Sci. 2023 May 5;24(9). doi: 10.3390/ijms24098307. Review. PubMed PMID: 37176013; PubMed Central PMCID: PMC10179602.
2. Righetto I, Gasparotto M, Casalino L, Vacca M, Filippini F. Exogenous Players in Mitochondria-Related CNS Disorders: Viral Pathogens and Unbalanced Microbiota in the Gut-Brain Axis. 2023 Jan 13;13(1). doi: 10.3390/biom13010169. Review. PubMed PMID: 36671555; PubMed Central PMCID: PMC9855674.
3. Casalino L, Talotta F, Cimmino A, Verde P. The Fra-1/AP-1 Oncoprotein: From the “Undruggable” Transcription Factor to Therapeutic Targeting. Cancers (Basel). 2022 Mar 14;14(6). doi: 10.3390/cancers14061480. Review. PubMed PMID: 35326630; PubMed Central PMCID: PMC8946526.
4. Casalino L, Verde P. Multifaceted Roles of DNA Methylation in Neoplastic Transformation, from Tumor Suppressors to EMT and Metastasis. Genes (Basel). 2020 Aug 12;11(8). doi: 10.3390/genes11080922. Review. PubMed PMID: 32806509; PubMed Central PMCID: PMC7463745.
5. Talotta F, Casalino L, Verde P. The nuclear oncoprotein Fra-1: a transcription factor knocking on therapeutic applications’ door. 2020 Jun;39(23):4491-4506. doi: 10.1038/s41388-020-1306-4. Epub 2020 May 8. Review. PubMed PMID: 32385348.
6. D’Aniello C, Cermola F, Palamidessi A, Wanderlingh LG, Gagliardi M, Migliaccio A, Varrone F, Casalino L, Matarazzo MR, De Cesare D, Scita G, Patriarca EJ, Minchiotti G. Collagen Prolyl Hydroxylation-Dependent Metabolic Perturbation Governs Epigenetic Remodeling and Mesenchymal Transition in Pluripotent and Cancer Cells. Cancer Res. 2019 Jul 1;79(13):3235-3250. doi: 10.1158/0008-5472.CAN-18-2070. Epub 2019 May 6. PubMed PMID: 31061065.
7. D’Aniello C, Fico A, Casalino L, Guardiola O, Di Napoli G, Cermola F, De Cesare D, Tatè R, Cobellis G, Patriarca EJ, Minchiotti G. A novel autoregulatory loop between the Gcn2-Atf4 pathway and (L)-Proline metabolism controls stem cell identity. Cell Death Differ. 2015 Jul;22(7):1094-105. doi: 10.1038/cdd.2015.24. Epub 2015 Apr 10. PubMed PMID: 25857264; PubMed Central PMCID: PMC4572871.
8. Iannone M, Ventre M, Formisano L, Casalino L, Patriarca EJ, Netti PA. Nanoengineered surfaces for focal adhesion guidance trigger mesenchymal stem cell self-organization and tenogenesis. Nano Lett. 2015 Mar 11;15(3):1517-25. doi: 10.1021/nl503737k. Epub 2015 Feb 24.PubMed PMID: 25699511.
9. Casalino L, D’Ambra P, Guarracino MR, Irpino A, Maddalena L, Maiorano F, Minchiotti G, Patriarca EJ. Image Analysis and Classification for High-Throughput Screening of Embryonic Stem Cells. In: Zazzu V, Ferraro MB, Guarracino M, editors. Mathematical Models in Biology. Bringing Mathematics to Life; 2014 October 27; Naples, NA, ITALY. DOI 10.1007/978-3-319-23497-7 Print ISBN 978-3-319-23496-0 Online ISBN 978-3-319-23497-7: Springer International Publishing; c2015.

Actual position:

CNR Research Scientist, since February 2010

Professional Roles:

Since Jan 2025: Coordinator of the OASI Research Biobank for Asplenia

Since 2020: Principal Investigator

at the IGB-ABT, CNR of Naples

Developmental Biology and Genetics

April 2006 – 2020 Responsible for the Molecular Screening Laboratory

“Stem Cell Fate Lab”, at the IGB-ABT, CNR of Naples.

“An automated robotic platform designed to identify molecules that control self-renewal, pluripotency, and differentiation of stem cells”.

April 2006 – January 2010 Research Fellow

Supervisor: Dr. Eduardo J. Patriarca

“Stem Cell Fate Lab”, at the IGB-ABT, CNR of Naples.

“The role of L-Pro in metabolic control of mESC metastability and pluripotency.”

June 2004 – March 2006: Post-doctoral fellow

Supervisor: Prof. Moshe Yaniv

Department of Developmental Biology, Pasteur Institute, Paris, France.

“Fra-1 as a cell cycle sensor and regulator in thyroid neoplastic cells: the cyclin gene A identified as a new transcriptional target of Fra-1”.

2001-2003: Visiting Fellow

Supervisor: Prof. Moshe Yaniv

Department of Developmental Biology, Pasteur Institute, Paris, France.

“Molecular mechanisms controlling the proliferation of transformed thyroid cells”.

2000 – May 2004: Post-doctoral fellow

Supervisor: Dr. Pasquale Verde

Laboratory of Human Genetics, at the IGB-ABT of the CNR in Naples.

“The role of Fra-1 in the control of the cell cycle of thyroid neoplastic cells”.

1995 –1999: PhD student

Supervisor: Dr. Pasquale Verde

Laboratory of Human Genetics, at the IGB-ABT of the CNR in Naples.

“Positive and negative transcriptional control mechanisms in the human urokinase gene enhancer region”

“Role of the transcriptional factors AP-1 and NF-kB in the neoplastic transformation of thyroid cells”

January 1993 – March 1995: Graduate student

Supervisor: Prof. Francesco Blasi

Department of Genetics, University of Milan.

“The transcription factors AP-1 and NF-kB as mediators of the response to tumor promoters and human cytokines of the urokinase gene in lung cancer cells”.