The alliance between genetic biobanks and patient organisations: the experience of the telethon network of genetic biobanks

Background: Rare diseases (RDs) are often neglected because they affect a small percentage of the population (6-8 %), which makes research and development of new therapies challenging processes. Easy access to high-quality samples and associated clinical data is therefore a key prerequisite for biomedical research. In this context, Genetic Biobanks are critical to developing basic, translational and clinical research on RDs. The Telethon Network of Genetic Biobanks (TNGB) is aware of the importance of biobanking as a service for patients and has started a dialogue with RD-Patient Organisations via promotion of dedicated meetings and round-tables, as well as by including their representatives on the TNGB Advisory Board. This has enabled the active involvement of POs in drafting biobank policies and procedures, including those concerning ethical issues. Here, we report on our experience with RD-Patient Organisations who have requested the services of existing biobanks belonging to TNGB and describe how these relationships were established, formalised and maintained. Results: The process of patient engagement has proven to be successful both for lay members, who increased their understanding of the complex processes of biobanking, and for professionals, who gained awareness of the needs and expectations of the people involved. This collaboration has resulted in a real interest on the part of Patient Organisations in the biobanking service, which has led to 13 written agreements designed to formalise this process. These agreements enabled the centralisation of rare genetic disease biospecimens and their related data, thus making them available to the scientific community. Conclusions: The TNGB experience has proven to be an example of good practice with regard to patient engagement in biobanking and may serve as a model of collaboration between disease-oriented Biobanks and Patient Organisations. Such collaboration serves to enhance awareness and trust and to encourage the scientific community to address research on RDs

Saving temporary exhibitions in virtual environments: The Digital Renaissance of Ulisse Aldrovandi – Acquisition and digitisation of cultural heritage objects

As per the objectives of Project CHANGES, particularly its thematic sub-project on the use of virtual technologies for museums and art collections, our goal was to obtain a digital twin of the temporary exhibition on Ulisse Aldrovandi called “The Other Renaissance”, and make it accessible to users online. After a preliminary study of the exhibition, focusing on acquisition constraints and related solutions, we proceeded with the digital twin creation by acquiring, processing, modelling, optimising, exporting, and metadating the exhibition. We made hybrid use of two acquisition techniques to create new digital cultural heritage objects and environments, and we used open technologies, formats, and protocols to make available the final digital product. Here, we describe the process of collecting and curating bibliographical exhibition (meta) data and the beginning of the digital twin creation to foster its findability, accessibility, interoperability, and reusability. The creation of the digital twin is currently ongoing

Hemophagocytic inflammatory syndrome in ADA-SCID: report of two cases and literature review

Hemophagocytic inflammatory syndrome (HIS) is a rare form of secondary hemophagocytic lymphohistiocytosis caused by an impaired equilibrium between natural killer and cytotoxic T-cell activity, evolving in hypercytokinemia and multiorgan failure. In the context of inborn errors of immunity, HIS occurrence has been reported in severe combined immunodeficiency (SCID) patients, including two cases of adenosine deaminase deficient-SCID (ADA-SCID). Here we describe two additional pediatric cases of ADA-SCID patients who developed HIS. In the first case, HIS was triggered by infectious complications while the patient was on enzyme replacement therapy; the patient was treated with high-dose corticosteroids and intravenous immunoglobulins with HIS remission. However, the patient required HLA-identical sibling donor hematopoietic stem cell transplantation (HSCT) for a definitive cure of ADA-SCID, without HIS relapse up to 13 years after HSCT. The second patient presented HIS 2 years after hematopoietic stem cell gene therapy (GT), secondarily to Varicella-Zoster vaccination and despite CD4+ and CD8+ lymphocytes’ reconstitution in line with other ADA SCID patients treated with GT. The child responded to trilinear immunosuppressive therapy (corticosteroids, Cyclosporine A, Anakinra). We observed the persistence of gene-corrected cells up to 5 years post-GT, without HIS relapse. These new cases of children with HIS, together with those reported in the literature, support the hypothesis that a major dysregulation in the immune system can occur in ADA-SCID patients. Our cases show that early identification of the disease is imperative and that a variable degree of immunosuppression could be an effective treatment while allogeneic HSCT is required only in cases of refractoriness. A deeper knowledge of immunologic patterns contributing to HIS pathogenesis in ADA-SCID patients is desirable, to identify new targeted treatments and ensure patients’ long-term recovery

Disease Evolution and Response to Rapamycin in Activated Phosphoinositide 3-Kinase δ Syndrome: The European Society for Immunodeficiencies-Activated Phosphoinositide 3-Kinase δ Syndrome Registry

Activated phosphoinositide 3-kinase (PI3K) δ Syndrome (APDS), caused by autosomal dominant mutations in PIK3CD (APDS1) or PIK3R1 (APDS2), is a heterogeneous primary immunodeficiency. While initial cohort-descriptions summarized the spectrum of clinical and immunological manifestations, questions about long-term disease evolution and response to therapy remain. The prospective European Society for Immunodeficiencies (ESID)-APDS registry aims to characterize the disease course, identify outcome predictors, and evaluate treatment responses. So far, 77 patients have been recruited (51 APDS1, 26 APDS2). Analysis of disease evolution in the first 68 patients pinpoints the early occurrence of recurrent respiratory infections followed by chronic lymphoproliferation, gastrointestinal manifestations, and cytopenias. Although most manifestations occur by age 15, adult-onset and asymptomatic courses were documented. Bronchiectasis was observed in 24/40 APDS1 patients who received a CT-scan compared with 4/15 APDS2 patients. By age 20, half of the patients had received at least one immunosuppressant, but 2–3 lines of immunosuppressive therapy were not unusual before age 10. Response to rapamycin was rated by physician visual analog scale as good in 10, moderate in 9, and poor in 7. Lymphoproliferation showed the best response (8 complete, 11 partial, 6 no remission), while bowel inflammation (3 complete, 3 partial, 9 no remission) and cytopenia (3 complete, 2 partial, 9 no remission) responded less well. Hence, non-lymphoproliferative manifestations should be a key target for novel therapies. This report from the ESID-APDS registry provides comprehensive baseline documentation for a growing cohort that will be followed prospectively to establish prognostic factors and identify patients for treatment studies

Ten-year single center experience with semi-eversion carotid endarterectomy: analysis of a cohort of patients with tips and tricks of our technique

The two most common surgical techniques to prevent ischemic stroke in patients with asymptomatic significant extracranial atherosclerotic carotid disease, or in symptomatic patients, are conventional CEA with patch closure (C-CEA) and eversion CEA (E-CEA) (1,2). We describe and present our ten-year experience on the efficacy and safety of semi-eversion CEA (SE-CEA) technique. CEA was offered to 1273 patients for significant (>70%, according to NASCET criteria), or symptomatic, primitive carotid artery stenosis during 2008-2018 (3). All had a preoperative carotid duplex ultrasound scan (DUS). Few patients with DUS of difficult interpretation received a preoperative computed tomography or magnetic resonance angiography. Long-term surveillance was based on carotid DUS at 3, 6 months, and each year thereafter. Restenosis was defined as a reduction in the diameter of the target vessel of at least 70%, diagnosed by a peak systolic velocity of at least 3 m/sec (4). All patients received prior informed consent from the operator or assistant, in compliance to the Helsinki informed consent declaration. Surgery is performed under general anesthesia and near-infrared spectroscopy (NIRS) for cerebral monitoring of regional oxygen saturation (rSO2). Surgical exposure is achieved in the usual fashion. Unfractionated heparin (2500 IU) are administered to the patient. The common carotid artery (CCA) is carefully exposed and a silastic vessel loop is double passed around it. The dissection is continued distally to the carotid bifurcation, where a local injection of a few ml of lidocaine is often required to avoid bradycardia. The hypoglossal nerve is recognized and respected, and the dissection is continued to the ICA, external carotid artery (ECA), and superior thyroid artery (STA). Sufficient distal dissection of the ICA past the finger-palpable end of the plaque is required to allow clamping of the artery on the free-plaque portion. A silastic vessel loop is double passed around the ICA and ECA. Vascular clamps are applied to CCA, ECA and STA, and 1 minute neurological assessment is performed. If rSO2 decreased to < 20% of the baseline value, the procedure is converted to C-CEA with patch closure, and a Pruitt carotid shunt (LeMaitre Vascular, Inc) is inserted and open. If no rSO2 decrease occurs after clamping, a longitudinal arteriotomy of the postero-lateral wall of the carotid bifurcation, slightly extended to ICA, is performed (Figure 1). After checking its back-flow, a vascular clamp is applied to ICA. Then, with the aid of a Watson-Cheyne dissector, the CCA, carotid bifurcation, and ICA are dissected away from the plaque. The plaque is completely divided at the level of CCA (Figure 2). The plaque in the ECA is broken with the help of the vascular clamp, distal to its origin, and then pulled out of the artery with the help of a surgical forceps. With the aid of the dissector, the distal ICA is carefully dissected away from the distal part of the plaque, which is cranially followed up to its distal endpoint. The wall of the ICA is distally everted by the second operator with two forceps, and the plaque is removed in one piece by the operator with the surgical forceps (Figure 3); any residual intimal debris can be removed under direct vision before replacing the ICA to its normal configuration. If the operator is not satisfied with his visualization of the endpoint of the plaque, the arteriotomy is extended distally along the ICA, and a C-CEA with patch closure and a Pruitt carotid shunt (LeMaitre Vascular, Inc) is employed. It is important to create a smooth dissection between the arterial wall and the plaque to limit the development of intimal flaps and debris, allowing the one piece-plaque removal. After checking the back-flow from the ICA and moving the clamp cranially, the arteriotomy is primarily closed with a continuous 6-0 Premio (Peters Surgical International Co, Ltd) round trip suture (Figure 4). Right before ending the suture, flushes from all carotid vessels are performed. The clamps are then removed from the ECA and CCA, and after some seconds from the ICA. The operating time is concluded in the usual fashion. Demographic, clinical, surgical, outcome variables, and follow-up of the entire cohort are presented in the Table. The EVEREST multicenter randomized trial compared C-CEA and E-CEA (3), finding no significant difference in postoperative mortality rate or in the onset of new neurological deficit between these two techniques, and demonstrating that C-CEA requires shorter clamp times than E-CEA and, when a patch was used during C-CEA, both techniques had a similar low rate of restenosis. We demonstrated a low ICA restenosis rate at a mean follow-up of 38 mo (the mean time of restenosis detection was 12 mo), comparable to any CEA restenosis rate reported in a recent systematic review (5). With SE-CEA a low restenosis rate may be ensured by the surgeon’s skill in performing the arteriotomy. Indeed, when performing the arteriotomy on the lateral-posterior wall of the carotid bifurcation, slightly extended to ICA, it is possible to suture the carotid in a wide area of the arterial wall itself, avoiding to incur in suture defects. Furthermore, by removing the plaque from ICA under the direct view of the endpoint it is unlikely leaving out the ending part of the plaque, causing residual stenosis. In our experience, patients treated with SE-CEA present a perioperative stroke and death rates widely less than 3%. Despite the encouraging results, our technique presents some limitations. In case of very long plaque it may be difficult to clearly define an endpoint into the ICA, and SE-CEA proves to be impractical. Conversion to C-CEA with patch closure is recommended in these cases, and we prefer to shunt when using a patch. Furthermore, in cases of shunt required for rSO2 decrease to less than 20% of the baseline value, we prefer to extent the arteriotomy distally into the ICA, for more safety when inserting the Pruitt shunt, hence performing a conversion to patch closure. This may represent another limitation of our technique. However, this scenario occurred only in few cases, indeed the most critical patients (i.e., those with occlusion of the contralateral ICA, and those with significant decrease of cerebral rSO2 detected by NIRS during carotid cross-clamping)

Autoantibodies to Harmonin and Villin Are Diagnostic Markers in Children with IPEX Syndrome

<div><p>Autoantibodies to enterocyte antigens harmonin (75 kDa USH1C protein) and villin (actin-binding 95 kDa protein) are associated with the Immune dysregulation, Polyendocrinopathy, Enteropathy, X-linked (IPEX) syndrome. In this study we evaluated the diagnostic value of harmonin and villin autoantibodies in IPEX and IPEX-like syndromes. Harmonin and villin autoantibodies were measured by a novel Luminescent-Immuno-Precipitation-System (LIPS) quantitative assay, in patients with IPEX, IPEX-like syndrome, Primary Immunodeficiencies (PID) with enteropathy, all diagnosed by sequencing of the <i>FOXP3</i> gene, and in type 1 diabetes (T1D), celiac disease and healthy blood donors as control groups. Harmonin and villin autoantibodies were detected in 12 (92%) and 6 (46%) of 13 IPEX patients, and in none of the IPEX-like, PID, T1D, celiac patients, respectively. All IPEX patients, including one case with late and atypical clinical presentation, had either harmonin and/or villin autoantibodies and tested positive for enterocyte antibodies by indirect immunofluorescence. When measured in IPEX patients in remission after immunosuppressive therapy or hematopoietic stem cell transplantation, harmonin and villin autoantibodies became undetectable or persisted at low titers in all cases but one in whom harmonin autoantibodies remained constantly high. In one patient, a peak of harmonin antibodies paralleled a relapse phase of enteropathy. Our study demonstrates that harmonin and villin autoantibodies, measured by LIPS, are sensitive and specific markers of IPEX, differentiate IPEX, including atypical cases, from other early childhood disorders associated with enteropathy, and are useful for screening and clinical monitoring of affected children.</p></div

Scatter plot of HAA, VAA, and GADA titers in patients’ sera.

<p>HAA (panel A), VAA (panel B) and GADA (panel C) serum IgG titers expressed in arbitrary units in IPEX (n = 13), IPEX-like (n = 14), PID (n = 5), T1D (VAA and GADA n = 123, VAA n = 46), celiac disease patients (HAA n = 70, VAA n = 46, GADA n = 44), and in controls (HAA and VAA n = 123, GADA n = 67). Dotted line indicates the cut-off for positivity.</p

HAA and VAA titers in IPEX patients in the course of therapy.

<p>On the vertical axis are indicated HAA <b>(</b>diamonds<b>)</b> and VAA <b>(</b>triangles<b>)</b>, autoantibody titers expressed in arbitrary units, on the horizontal axis time in months. The vertical dotted line indicates the date of HSCT, horizontal dotted and dashed lines indicate the cut-off for positivity of HAA and VAA, respectively.</p

Immunofluorescent staining of intestinal enterocytes with patients’ sera.

<p>HAA from IPEX Pt 19 bind the brush border and cytosol of enterocytes (panel A) while VAA from IPEX Pt 17 binds only the brush border (panel B). IgG from PID Pt L1 bind the enterocytes brush border (panel C). Absence of binding in IPEX-like Pt L30 (panel D).</p