Maternal Folate Intake and Risk of Childhood Brain and Spinal Cord Tumors: A Systematic Review and Meta-Analysis

Background: Many epidemiological studies have investigated the effect of maternal diet and prenatal multivitamin supplementation on pediatric cancer risk. Childhood brain and spinal cord tumors (CBSCT) have been attributed to different possible risk factors. Methods: We conducted a systematic review and meta-analysis on maternal folate intake before and during pregnancy and the risk of CBSCT. We systematically reviewed publications obtained by searching the Insitute for Scientific Information Web of Knowledge and PubMed literature databases. We extracted the risk estimate of the highest and the lowest reported categories of intake from each study and conducted a meta-analysis using a random-effects model. Results: The results of the pooled analysis of all 10 studies, 1 cohort and 9 case-control studies, indicated that maternal folate intake was inversely associated with CBSCT risk (OR 0.77; 95% CI 0.67–0.88, p < 0.001; I2 = 51.22%, p = 0.001). Separate analyses on the basis of the source of folate (folic acid supplementation, dietary folate) and in relation to the timing of exposure (before pregnancy, during pregnancy) found that folic acid supplementation was associated with an approximately 23% reduction in ­CBSCT risk (OR 0.77, 95% CI 0.66–0.90, p = 0.001; I2 = 53.18%, p = 0.001) and consumption during pregnancy was associated with an approximately 20% reduction in CBSCT risk (OR 0.80, 95% CI 0.67–0.97, p = 0.020; I2 = 62.48%, p < 0·001). Conclusions: Maternal consumption of folic acid is associated with a reduced risk of CBSCT. Further investigations are necessary to increase the reliability of the results and estimate the relationship between dose-response and the best outcome

How to promote changes in primary care? The Florentine experience of the House of Community

Primary care (PC) has a central role in promoting health and preventing diseases, even during health emergencies. The COVID-19 pandemic has shown how strengthening comprehensive primary healthcare (c-PHC) services is key to ensuring community health. The Italian government decided to support PHC by investing resources from the Next Generation EU (NextGenEu) plan in the development of local health districts (LHDs) and local PC centers called “Houses of Community (HoC)”. The Florence LHD (Tuscany)—in direct collaboration with the University of Florence—has represented the experimental context in which a c-PHC-inspired organizational model has been proposed and included the HoC as the nearest access point to PC services. Through multiprofessional collaboration practices, HoCs provide continuity of care as well as health and social integration. Different levels of action must coexist to initiate, implement, and sustain this new PC model: the organizational and managerial level, the experimentation of a new model of care, and the research level, which includes universities and LHD through participatory research and action approaches. This process benefits from health professionals' (HPs) participation and continuous assessment, the care for working relationships between HPs and services, an appropriate research methodology together with a “permeable” multidisciplinary research group, and educational programs. In this context, the HoC assumes the role of a permanent laboratory of experimentation in PC, supporting the effectiveness of care and answering what the Next Gen EU plan has been foreseeing for the rethinking of Italian territorial services

42 correction of scid x1 by targeted genome editing of hematopoietic stem progenitor cells hspc in the mouse model

Targeted genome editing by engineered nucleases has brought the goal of gene correction within the reach of gene therapy. A candidate disease for HSPC gene correction is SCID-X1, because gene therapy trials with integrating vectors showed robust clinical efficacy even from few corrected cells but also the occurrence of leukemias due to insertional mutagenesis and unregulated transgene expression. To model SCID-X1 gene correction in preclinical studies, we developed a mouse model carrying the IL2RG human gene harboring a common disease-causing mutation in place of the murine Il2rg, allowing to use of the same reagents developed for gene correction of human cells. These mice have impaired lymphoid development which phenocopies that reported for Il2rg-/- mice. To assess the minimal level of corrected HSPC required to achieve immune reconstitution we performed competitive transplants with wild-type (WT) and Il2rg-/- HSPC and found that 1% of WT cells are sufficient to reconstitute in part the T and B cell compartments. We then tested gene correction of the murine Lin- HSPC by the delivery of donor DNA template by IDLVs followed by transfection of ZFN mRNAs. This protocol yielded high on-target nuclease activity (40%) and a mean of 6% transgene integration by HDR but also high cytotoxicity (65% cell loss) under the conditions we used. The surviving cells remained capable of expansion in culture and maintained their clonogenic potential. Importantly, upon transplant into lethally irradiated mice, only the gene corrected cells were able to generate lymphoid lineages (B and T cells), showing a clear selective advantage over the un-corrected SCID cells. These data indicate functional correction of the defective IL2RG gene by targeted editing. Furthermore, upon challenging the mice with a murine pathogen we observed viral-specific γIFN production by CD8+ gene corrected cells, proving their in vivo functionality. Yet, measuring the percentage of edited cells (either by NHEJ or HDR) within the HSC compartment long-term, we found that it was nearly undetectable. Despite the lack of HSC marking, gene corrected lymphoid cells persisted in the mice up to 7 months post transplantation within all the hematopoietic organs, indicating successful editing of at least 1% progenitors able to sustain long-term lymphopoiesis and partially correct the disease phenotype. We then developed a new protocol exploiting CRISPR/Cas9 technology that enabled to achieve substantial levels of targeted DNA repair by NHEJ (up to 70%) and HDR (up to 25%) with minimal cytotoxicity and provided stable engraftment of the edited cells in transplanted mice. By this strategy we are now assessing the impact of HSC vs. progenitor targeted editing and conditioning regimen for the extent and stability of disease correction. These studies will help establish the key factors underlying safe and effective rescue of the disease by HSPC gene editing and assist in the design of the protocol for its first clinical testing

481. Targeted Genome Editing in Mouse Hematopoietic Stem/Progenitor Cells (HSPC) To Model Gene Correction of SCID-X1

Targeted genome editing by artificial nucleases has brought the goal of gene correction within the reach of gene therapy. A candidate disease for HSPC gene correction is SCID-X1, because gene therapy with early generation integrating vectors showed robust clinical efficacy even from few corrected cells but also the occurrence of adverse events due to insertional mutagenesis and unregulated transgene expression. We recently reported a strategy that enabled targeted integration of a corrective cDNA into the IL2RG gene in 6% of human HSPC with high specificity. Gene corrected HSPC generated polyclonal lymphoid cells that express the IL2RG protein and have a selective growth advantage over those carrying disruptive IL2RG mutations (Genovese, Nature, 2014). Here, to model SCID-X1 disease correction, we developed a mouse model carrying the IL2RG human gene including a common disease-causing mutation in place of the murine Il2rg gene, allowing to use the same reagents utilized for gene correction of human cells. These mice have impairment in lymphoid development which phenocopies that reported for Il2rg-/- mice. To assess the minimal level of corrected HSPC required to achieve immune reconstitution we first performed competitive transplants with wild-type (WT) and Il2rg-/- HSPC and found that 1% of WT cells are sufficient to reconstitute at least in part the T and B cell compartments. We then developed a protocol to obtain gene correction in murine Lin- HSPC based on the delivery of donor DNA template by IDLVs followed by transfection of ZFN mRNAs. This protocol was associated with high on-target nuclease activity (40%) and a mean of 6% transgene integration by HDR, but also with high levels of acute cytotoxicity (65% cell loss). The surviving cells remained capable of expansion in culture and preserved their clonogenic potential. Importantly, upon transplant into lethally irradiated mice, only the gene corrected cells were able to generate lymphoid lineages (B and T cells), showing a clear selective advantage over un-corrected cells. These data indicate functional correction of the IL2RG gene by our strategy. Yet, measuring percentage of correction within myeloid cells at long-term we found that it was almost undetectable. Despite the lack of HSC marking, gene corrected lymphoid cells stably persisted in the mice up to 7 months post transplantation within all the hematopoietic organs. Furthermore, upon challenging the transplanted mice with a murine pathogen (LCMV Arm.) we observed viral-specific γIFN production by CD8+ gene corrected cells at a similar extent as for WT mice, proving in vivo the functionality of corrected T cells. These results suggest that our protocol achieves biologically relevant levels of gene correction in progenitors that sustain long-term lymphopoiesis but is limited in multipotent HSC. Ongoing studies aim to improve murine HSC gene targeting and to compare safety and efficacy of gene correction vs gene replacement in our disease model

The Activity of Special Continuity Care Units in the City of Florence During the COVID-19 Pandemic

Objectives: Worldwide, countries adopted different strategies in primary care (PC) to cope with the COVID-19 pandemic. This study aims to describe and evaluate the functions and activity load of a specific PC organizational model called “Special Continuity Care Units” (SCCU) in Florence, Italy, and to investigate the characteristics of the COVID-19 patients assisted by the service.Methods: The retrospective cross-sectional design used daily updated reports by SCCU team members to evaluate the activity load. The retrospective cohort study analyzed data of the demographics, clinical characteristics, and process outcomes of patients assisted during the second pandemic wave.Results: The analysis shows how the service activity load changed along with the epidemiological trend. Regarding people assisted by the SCCU, the median follow-up duration of symptoms was 6 days; male gender and being symptomatic were predictors of hospitalization.Conclusion: Some key characteristics can be described as indispensable in PC services facing health emergencies: model flexibility, the availability of resources, networking among services to enhance coordination and resource optimization, and close collaboration with general practitioners

現地観測に基づく河川流の乱流特性に関する研究

博士（工学）神戸大

Engineered tumor-infiltrating macrophages as gene delivery vehicles for interferon-α activates immunity and inhibits breast cancer progression

An immunosuppressive tumor microenvironment is a cancer hallmark and a major impediment to successful immunotherapy. We engineered hematopoietic progenitors to target expression of an interferon-α (IFNα) transgene specifically to their monocytic progeny, including tumor-infiltrating macrophages. Mice chimeric for these IFNα-expressing macrophages showed activation of innate and adaptive immune cells against breast cancer and inhibited disease progression

Exogenous Hormone Factors in Relation to the Risk of Malignant Melanoma in Women: A Systematic Review and Meta-Analysis

The influence of exogenous female hormones on the risk of developing malignant melanoma in women remains controversial. The aim of our review and meta-analysis is to summarize the evidence and derive a more accurate estimation of the association between oral contraceptives (OCs) or menopausal hormone therapy (MHT) and the risk of developing malignant melanoma in women. PubMed, Web of Science, and Scopus database were searched for studies published up until October 2021. The PRISMA statement and MOOSE guidelines were followed. Studies were pooled using a random effects model. Heterogeneity was explored with the chi-square-based Cochran’s Q statistic and the I2 statistic. Publication bias was assessed with Begg’s test and Egger’s test. Forty-six studies met the eligibility criteria. The pooled analysis (26 studies) on OC use and the risk of developing cutaneous malignant melanoma (CMM) showed no significant association, but demonstrated significant association for cohort studies (OR 1.08, 95% CI 1.01–1.16; I2 = 0.00%, p = 0.544). The pooled analysis (16 studies) showed a significantly increased risk of CMM in association with MHT (OR 1.15, 95% CI 1.08–1.23; I2 = 25.32%, p = 0.169). Stratifying the results by study design showed that a significant increased risk of CMM was associated with MHT in the cohort studies (OR 1.12; 95% CI 1.04–1.19; I2 = 0%, p = 0.467). No significant publication bias could be detected. Further studies are needed to investigate the potential association with formulation, duration of use, and dosage of use, and to better understand the role of possible confounders