Cellular and Molecular Characterization of Breast Cancer Chemotherapy-Related B Cell Lymphomas

Chemotherapy can prolong life expectancy in patients with hematologic malignancies. However, the long-lasting effects of chemotherapy on the function of hematopoietic and immune systems remain to be fully characterized. The hematopoietic and immune systems are vital for lifelong supply of blood cells and defense against pathogens and malignancies. Cyclophosphamide (CTX) and Doxorubicin (DOX) are frequently used together as neoadjuvant and adjuvant therapy for breast cancer. The main goal of our project is to characterize in depth long-term adverse effects of CTX/DOX treatment on the function of hematopoietic and immune systems. A cohort of Ly5.2+ C57BL6 female mice were treated once a week for 5 consecutive weeks with 30 mg/kg CTX and 3 mg/kg DOX. Cohorts of age-matched control mice and CTX/DOX-treated mice were analyzed 15 months after the end of treatment. Comprehensive flow cytometry analysis of >50 hematopoietic and immune cell types (HSPCs, B cells, T cells, myeloid, erythroid and megakaryocytic lineages) from the bone marrow (BM) and spleen (SPL) of individual age-matched control and CTX/DOX-treated mice, in combination with BM and SPL cell transplant from each mouse has revealed development of CTX/DOX-related B cell lymphoma (BCL) in 17% of CTX/DOX-treated mice. Moreover, 13% of the CTX/DOX-treated mice showed early stages of BCL development, while the remaining 70% of CTX/DOX-treated and all age-matched control mice did not show any signs of BCL. Chemo-treated mice developing BCL exhibited expansion of lymphoid-biased short-term HSCs (ST-HSC), significant reduction of Long-term HSCs (LT-HSC) in BM and SPL and increased population of immature B cell in the SPL. Transplant of BM and SPL cells from CTX/DOX-treated lymphoma-bearing (CTX/DOX-BCL) mice into Ly5.1+ syngeneic recipient mice resulted in massive splenomegaly, expansion of Ly5.2+ lymphoid-biased ST-HSCs and CLPs in the SPL, reduced Ly5.2+ myeloid-biased HSCs in BM and SPL and reduced Ly5.2+ MPPs and CMPs cells in the BM. Additionally, all Ly5.2+ B cell developmental stages were increased in CTX/DOX-BCL recipients. B1 cells were also significantly increased in the lymphoma transplant recipient mice, mimicking B1 cell overproduction in their donors. These results indicate that CTX/DOX treatment increases the incidence of cancer therapy-related B cell lymphoma development. Further immunophenotypic characterization of donor Ly5.2+ B cells in recipients identified coexistence of two BCL types: 1) CLL/SLL (CD19+ CD23+ CD20+ CD5+ B220-), and 2) Mantle Cell Lymphoma (MCL; CD19+ CD23- CD20+ CD5+ B220-), where MCL is predominant in BM and SPL. In contrast, mice transplanted with cells from CTX/DOX-treated "early stage lymphoma" mice did not have exorbitant amount of these two immunophenotypic populations in the SPL as compared to CTX/DOX-BCL recipient mice; but BM had a 2-fold increase of these cells compared to control. Likewise, transplant recipients of cells from "early stage lymphoma" and "normal" CTX/DOX donor mice had increased number of lymphoid-biased ST-HSCs in the SPL and reduced LT-HSCs in BM. Adding to the adverse effects of CTX/DOX on the immune system, populations of Natural Killer (NK) and Natural Killer T (NKT) cells were significantly decreased in recipients of cells from all CTX/DOX-treated mice compared to control transplant recipients. More importantly, RNAseq analysis revealed: 1) reduced expression of NK receptors (NKG2D, NKp46, CD94, CD69), 2) downregulated expression of PI3K, Fyn and PLCγ genes involved in NK cell-mediated cytotoxicity, and 3) upregulation of BAG6 and ULBP1 genes, known as suppressors of NK receptors, in CTX/DOX-BCL recipients compared to control recipients. Combined, these results suggest that attenuated numbers and impaired function of NK/NKT cells could be accountable for development of chemotherapy-related B cell lymphomas. Dysregulation of the hematopoietic system, expansion of BCL populations and attenuated NK/NKT function/surveillance in CTX/DOX-treated mice and mice transplanted with CTX/DOX-BCL cells demonstrate some of the harmful effects of CTX/DOX on the hematopoietic and immune system, causing the co-development of CLL and MCL B cell lymphomas. Thus, it is important to investigate direct effects of CTX/DOX on NK/NKT cells and determine the molecular and cellular causes and cellular origin (HSC, Ly-HSC, CLPs, B1 cells) of cancer therapy-related B cell lymphomas. Disclosures No relevant conflicts of interest to declare

Neoantigen-specific CD4+ tumor-infiltrating lymphocytes are potent effectors identified within adoptive cell therapy products for metastatic melanoma patients

Background Adoptive cell therapy (ACT) with tumor-infiltrating lymphocytes (TILs) is a promising immunotherapeutic approach for patients with advanced solid tumors. While numerous advances have been made, the contribution of neoantigen-specific CD4+T cells within TIL infusion products remains underexplored and therefore offers a significant opportunity for progress.Methods We analyzed infused TIL products from metastatic melanoma patients previously treated with ACT for the presence of neoantigen-specific T cells. TILs were enriched on reactivity to neoantigen peptides derived and prioritized from patient sample-directed mutanome analysis. Enriched TILs were further investigated to establish the clonal neoantigen response with respect to function, transcriptomics, and persistence following ACT.Results We discovered that neoantigen-specific TIL clones were predominantly CD4+ T cells and were present in both therapeutic responders and non-responders. CD4+ TIL demonstrated an effector T cell response with cytotoxicity toward autologous tumor in a major histocompatibility complex class II-dependent manner. These results were validated by paired TCR and single cell RNA sequencing, which elucidated transcriptomic profiles distinct to neoantigen-specific CD4+ TIL.Conclusions Despite methods which often focus on CD8+T cells, our study supports the importance of prospective identification of neoantigen-specific CD4+ T cells within TIL products as they are a potent source of tumor-specific effectors. We further advocate for the inclusion of neoantigen-specific CD4+ TIL in future ACT protocols as a strategy to improve antitumor immunity

Adolescent transport and unintentional injuries: a systematic analysis using the Global Burden of Disease Study 2019

Background Globally, transport and unintentional injuries persist as leading preventable causes of mortality and morbidity for adolescents. We sought to report comprehensive trends in injury-related mortality and morbidity for adolescents aged 10-24 years during the past three decades. Methods Using the Global Burden of Disease, Injuries, and Risk Factors 2019 Study, we analysed mortality and disability-adjusted life-years (DALYs) attributed to transport and unintentional injuries for adolescents in 204 countries. Burden is reported in absolute numbers and age-standardised rates per 100 000 population by sex, age group (10-14, 15-19, and 20-24 years), and sociodemographic index (SDI) with 95% uncertainty intervals (UIs). We report percentage changes in deaths and DALYs between 1990 and 2019. Findings In 2019, 369 061 deaths (of which 214337 [58%] were transport related) and 31.1 million DALYs (of which 16.2 million [52%] were transport related) among adolescents aged 10-24 years were caused by transport and unintentional injuries combined. If compared with other causes, transport and unintentional injuries combined accounted for 25% of deaths and 14% of DALYs in 2019, and showed little improvement from 1990 when such injuries accounted for 26% of adolescent deaths and 17% of adolescent DALYs. Throughout adolescence, transport and unintentional injury fatality rates increased by age group. The unintentional injury burden was higher among males than females for all injury types, except for injuries related to fire, heat, and hot substances, or to adverse effects of medical treatment. From 1990 to 2019, global mortality rates declined by 34.4% (from 17.5 to 11.5 per 100 000) for transport injuries, and by 47.7% (from 15.9 to 8.3 per 100000) for unintentional injuries. However, in low-SDI nations the absolute number of deaths increased (by 80.5% to 42 774 for transport injuries and by 39.4% to 31 961 for unintentional injuries). In the high-SDI quintile in 2010-19, the rate per 100 000 of transport injury DALYs was reduced by 16.7%, from 838 in 2010 to 699 in 2019. This was a substantially slower pace of reduction compared with the 48.5% reduction between 1990 and 2010, from 1626 per 100 000 in 1990 to 838 per 100 000 in 2010. Between 2010 and 2019, the rate of unintentional injury DALYs per 100 000 also remained largely unchanged in high-SDI countries (555 in 2010 vs 554 in 2019; 0.2% reduction). The number and rate of adolescent deaths and DALYs owing to environmental heat and cold exposure increased for the high-SDI quintile during 2010-19. Interpretation As other causes of mortality are addressed, inadequate progress in reducing transport and unintentional injury mortality as a proportion of adolescent deaths becomes apparent. The relative shift in the burden of injury from high-SDI countries to low and low-middle-SDI countries necessitates focused action, including global donor, government, and industry investment in injury prevention. The persisting burden of DALYs related to transport and unintentional injuries indicates a need to prioritise innovative measures for the primary prevention of adolescent injury

Open data from the first and second observing runs of Advanced LIGO and Advanced Virgo

Advanced LIGO and Advanced Virgo are monitoring the sky and collecting gravitational-wave strain data with sufficient sensitivity to detect signals routinely. In this paper we describe the data recorded by these instruments during their first and second observing runs. The main data products are gravitational-wave strain time series sampled at 16384 Hz. The datasets that include this strain measurement can be freely accessed through the Gravitational Wave Open Science Center at http://gw-openscience.org, together with data-quality information essential for the analysis of LIGO and Virgo data, documentation, tutorials, and supporting software

Prospective observational cohort study on grading the severity of postoperative complications in global surgery research

Background The Clavien–Dindo classification is perhaps the most widely used approach for reporting postoperative complications in clinical trials. This system classifies complication severity by the treatment provided. However, it is unclear whether the Clavien–Dindo system can be used internationally in studies across differing healthcare systems in high- (HICs) and low- and middle-income countries (LMICs). Methods This was a secondary analysis of the International Surgical Outcomes Study (ISOS), a prospective observational cohort study of elective surgery in adults. Data collection occurred over a 7-day period. Severity of complications was graded using Clavien–Dindo and the simpler ISOS grading (mild, moderate or severe, based on guided investigator judgement). Severity grading was compared using the intraclass correlation coefficient (ICC). Data are presented as frequencies and ICC values (with 95 per cent c.i.). The analysis was stratified by income status of the country, comparing HICs with LMICs. Results A total of 44 814 patients were recruited from 474 hospitals in 27 countries (19 HICs and 8 LMICs). Some 7508 patients (16·8 per cent) experienced at least one postoperative complication, equivalent to 11 664 complications in total. Using the ISOS classification, 5504 of 11 664 complications (47·2 per cent) were graded as mild, 4244 (36·4 per cent) as moderate and 1916 (16·4 per cent) as severe. Using Clavien–Dindo, 6781 of 11 664 complications (58·1 per cent) were graded as I or II, 1740 (14·9 per cent) as III, 2408 (20·6 per cent) as IV and 735 (6·3 per cent) as V. Agreement between classification systems was poor overall (ICC 0·41, 95 per cent c.i. 0·20 to 0·55), and in LMICs (ICC 0·23, 0·05 to 0·38) and HICs (ICC 0·46, 0·25 to 0·59). Conclusion Caution is recommended when using a treatment approach to grade complications in global surgery studies, as this may introduce bias unintentionally

The surgical safety checklist and patient outcomes after surgery: a prospective observational cohort study, systematic review and meta-analysis

© 2017 British Journal of Anaesthesia Background: The surgical safety checklist is widely used to improve the quality of perioperative care. However, clinicians continue to debate the clinical effectiveness of this tool. Methods: Prospective analysis of data from the International Surgical Outcomes Study (ISOS), an international observational study of elective in-patient surgery, accompanied by a systematic review and meta-analysis of published literature. The exposure was surgical safety checklist use. The primary outcome was in-hospital mortality and the secondary outcome was postoperative complications. In the ISOS cohort, a multivariable multi-level generalized linear model was used to test associations. To further contextualise these findings, we included the results from the ISOS cohort in a meta-analysis. Results are reported as odds ratios (OR) with 95% confidence intervals. Results: We included 44 814 patients from 497 hospitals in 27 countries in the ISOS analysis. There were 40 245 (89.8%) patients exposed to the checklist, whilst 7508 (16.8%) sustained ≥1 postoperative complications and 207 (0.5%) died before hospital discharge. Checklist exposure was associated with reduced mortality [odds ratio (OR) 0.49 (0.32–0.77); P\u3c0.01], but no difference in complication rates [OR 1.02 (0.88–1.19); P=0.75]. In a systematic review, we screened 3732 records and identified 11 eligible studies of 453 292 patients including the ISOS cohort. Checklist exposure was associated with both reduced postoperative mortality [OR 0.75 (0.62–0.92); P\u3c0.01; I2=87%] and reduced complication rates [OR 0.73 (0.61–0.88); P\u3c0.01; I2=89%). Conclusions: Patients exposed to a surgical safety checklist experience better postoperative outcomes, but this could simply reflect wider quality of care in hospitals where checklist use is routine

Critical care admission following elective surgery was not associated with survival benefit: prospective analysis of data from 27 countries

This was an investigator initiated study funded by Nestle Health Sciences through an unrestricted research grant, and by a National Institute for Health Research (UK) Professorship held by RP. The study was sponsored by Queen Mary University of London