Evidence for B cell exhaustion in chronic graft-versus-host disease

Chronic graft-versus-host disease (cGvHD) remains a major complication of allogeneic hematopoietic stem cell transplantation (HSCT). A number of studies support a role for B cells in the pathogenesis of cGvHD. In this study, we report the presence of an expanded population of CD19+CD21− B cells with features of exhaustion in the peripheral blood of patients with cGvHD. CD21− B cells were significantly increased in patients with active cGvHD compared to patients without cGvHD and healthy controls (median 12.2 versus 2.12 versus 3%, respectively; p < 0.01). Compared with naïve (CD27−CD21+) and classical memory (CD27+CD21+) B cells, CD19+CD21− B cells in cGvHD were CD10 negative, CD27 negative and CD20hi, and exhibited features of exhaustion, including increased expression of multiple inhibitory receptors such as FCRL4, CD22, CD85J, and altered expression of chemokine and adhesion molecules such as CD11c, CXCR3, CCR7, and CD62L. Moreover, CD21− B cells in cGvHD patients were functionally exhausted and displayed poor proliferative response and calcium mobilization in response to B-cell receptor triggering and CD40 ligation. Finally, the frequencies of circulating CD21− B cells correlated with cGvHD severity in patients after HSCT. Our study further characterizes B cells in chronic cGVHD and supports the use of CD21−CD27−CD10− B cell frequencies as a biomarker of disease severity

Combined inhibition of BCL-2 and MCL-1 overcomes BAX deficiency-mediated resistance of TP53-mutant acute myeloid leukemia to individual BH3 mimetics

TP53-mutant acute myeloid leukemia (AML) respond poorly to currently available treatments, including venetoclax-based drug combinations and pose a major therapeutic challenge. Analyses of RNA sequencing and reverse phase protein array datasets revealed significantly lower BAX RNA and protein levels in TP53-mutant compared to TP53-wild-type (WT) AML, a finding confirmed in isogenic CRISPR-generated TP53-knockout and -mutant AML. The response to either BCL-2 (venetoclax) or MCL-1 (AMG176) inhibition was BAX-dependent and much reduced in TP53-mutant compared to TP53-WT cells, while the combination of two BH3 mimetics effectively activated BAX, circumventing survival mechanisms in cells treated with either BH3 mimetic, and synergistically induced cell death in TP53-mutant AML and stem/progenitor cells. The BH3 mimetic-driven stress response and cell death patterns after dual inhibition were largely independent of TP53 status and affected by apoptosis induction. Co-targeting, but not individual targeting of BCL-2 and MCL-1 in mice xenografted with TP53-WT and TP53-R248W Molm13 cells suppressed both TP53-WT and TP53-mutant cell growth and significantly prolonged survival. Our results demonstrate that co-targeting BCL-2 and MCL-1 overcomes BAX deficiency-mediated resistance to individual BH3 mimetics in TP53-mutant cells, thus shifting cell fate from survival to death in TP53-deficient and -mutant AML. This concept warrants clinical evaluation

Combined Inhibition of Bcl-2 and Mcl-1 Overcomes Bax Deficiency-Mediated Resistance of TP53-Mutant Acute Myeloid Leukemia to Individual BH3 Mimetics

TP53-mutant acute myeloid leukemia (AML) respond poorly to currently available treatments, including venetoclax-based drug combinations and pose a major therapeutic challenge. Analyses of RNA sequencing and reverse phase protein array datasets revealed significantly lower BAX RNA and protein levels in TP53-mutant compared to TP53-wild-type (WT) AML, a finding confirmed in isogenic CRISPR-generated TP53-knockout and -mutant AML. The response to either BCL-2 (venetoclax) or MCL-1 (AMG176) inhibition was BAX-dependent and much reduced in TP53-mutant compared to TP53-WT cells, while the combination of two BH3 mimetics effectively activated BAX, circumventing survival mechanisms in cells treated with either BH3 mimetic, and synergistically induced cell death in TP53-mutant AML and stem/progenitor cells. The BH3 mimetic-driven stress response and cell death patterns after dual inhibition were largely independent of TP53 status and affected by apoptosis induction. Co-targeting, but not individual targeting of BCL-2 and MCL-1 in mice xenografted with TP53-WT and TP53-R248W Molm13 cells suppressed both TP53-WT and TP53-mutant cell growth and significantly prolonged survival. Our results demonstrate that co-targeting BCL-2 and MCL-1 overcomes BAX deficiency-mediated resistance to individual BH3 mimetics in TP53-mutant cells, thus shifting cell fate from survival to death in TP53-deficient and -mutant AML. This concept warrants clinical evaluation

Detection of subclinical keratoconus using biometric parameters

The validation of innovative methodologies for diagnosing keratoconus in its earliest stages is of major interest in ophthalmology. So far, subclinical keratoconus diagnosis has been made by combining several clinical criteria that allowed the definition of indices and decision trees, which proved to be valuable diagnostic tools. However, further improvements need to be made in order to reduce the risk of ectasia in patients who undergo corneal refractive surgery. The purpose of this work is to report a new subclinical keratoconus detection method based in the analysis of certain biometric parameters extracted from a custom 3D corneal model. This retrospective study includes two groups: the first composed of 67 patients with healthy eyes and normal vision, and the second composed of 24 patients with subclinical keratoconus and normal vision as well. The proposed detection method generates a 3D custom corneal model using computer-aided graphic design (CAGD) tools and corneal surfaces’ data provided by a corneal tomographer. Defined bio-geometric parameters are then derived from the model, and statistically analysed to detect any minimal corneal deformation. The metric which showed the highest area under the receiver-operator curve (ROC) was the posterior apex deviation. This new method detected differences between healthy and sub-clinical keratoconus corneas by using abnormal corneal topography and normal spectacle corrected vision, enabling an integrated tool that facilitates an easier diagnosis and follow-up of keratoconus.This publication has been carried out in the framework of the Thematic Network for Co-Operative Research in Health (RETICS) reference number RD16/0008/0012 financed by the Carlos III Health Institute-General Subdirection of Networks and Cooperative Investigation Centers (R&D&I National Plan 2013–2016) and the European Regional Development Fund (FEDER)

Protective Policy Index (PPI) global dataset of origins and stringency of COVID 19 mitigation policies

This the final version. Available on open access from Nature Research via the DOI in this recordData Records: We have created a Github repository (https://github.com/COVID-policy-response-lab/PPI-data) to store the datasets with the Public Health Protective Policy Index and its components. A copy of the included datafiles, as described below, was deposited with openICPSR15. It presently requires creating an account with the depository. Data access is free. Data location is at https://www.openicpsr.org/openicpsr/project/123401. The datasets are stored as csv files with five types of layouts. “PPI_country_m1.csv” is a file with country-level aggregates of region-level PPIs, computed using method 1, and their components. Each row corresponds to a country-date. The rows are identified using the country name (cname), numeric and 2-letter ISO 3166-1 codes (isocode and isoabbr respectively), as well as a date variable. The names of the policy variables contain four components: the name of the broader category, the name of the category, the level of issuing government (“nat” refers to the national policies, “reg” refers to the subnational policies, and “tot” refers to the combination of national and subnational policies), as well as suffix “ave”. For example, the average Total PPI is denoted as “ppi.all.tot.ave”, and the average stringency of the closures of air borders by the national government is denoted as “borders.air_bord.nat.ave”. See the codebook for the complete list of variables. “PPI_country_m2.csv” is a file with country-level aggregates of region-level PPIs, computed using method 2, and their components. The identifying variables and the naming convention for the policy variables is the same as in “PPI_country_m1.csv”, with the addition of suffix “0.2” at the end of the policy variable names. “PPI_regions_XX_m1.csv” (replace XX with the 2-letter ISO 3166-1 country codes) are country-specific files with region-specific PPIs, computed using method 1, and their components. The identifying variables include the numeric and 2-letter ISO 3166-1 codes of the country (isocode and isoabbr respectively), the name of the region (state_province), its ISO 3166-2 code (iso_state), as well as a date variable. The names of the policy variables contain three components: the name of the broader category, the name of the category, and the level of issuing government (“nat” refers to the national policies, “reg” refers to the subnational policies, and “tot” refers to the combination of national and subnational policies). For example, the average Total PPI is denoted as “ppi.all.tot”, and the stringency of the closures of air borders by the national government is denoted as “borders.air_bord.nat”. “PPI_regions_XX_m2.csv” (replace XX with the 2-letter ISO 3166-1 country codes are country-specific files with region-specific PPIs, computed using method 2, and their components. The identifying variables and the naming convention for the policy variables is the same as in “PPI_regions_XX_m1.csv”, with the addition of the suffix “0.2” at the end of the policy variable names. “changes_regions_m1.csv” is an auxiliary file that describes the changes in the policy states, as recorded in the “PPI_regions_XX_m1.csv” files. Each row in this file corresponds to a change in a value of a policy state variable in a region and of a specific government level. The case identifying variables include the name of the country (cname), the numeric and 2-letter ISO 3166-1 code of the country (isocode and isoabbr, respectively), the name of the region (state_province) and its ISO 3166-2 code, date, policy dimension, and a marker of policies issued by a regional government (subnational). Among others, the attributes included in this file include the branch of the government (branch) and the date when the change was announced (report_date).Code availability; The code used to produce our calculations is available at https://github.com/COVID-policy-response-lab/PPI-dataWe have developed and made accessible for multidisciplinary audience a unique global dataset of the behavior of political actors during the COVID-19 pandemic as measured by their policy-making efforts to protect their publics. The dataset presents consistently coded cross-national data at subnational and national levels on the daily level of stringency of public health policies by level of government overall and within specific policy categories, and reports branches of government that adopted these policies. The data on these public mandates of protective behaviors is collected from media announcements and government publications. The dataset allows comparisons of governments’ policy efforts and timing across the world and can serve as a source of information on policy determinants of pandemic outcomes–both societal and possibly medical

Cord blood NK cells engineered to express IL-15 and a CD19-targeted CAR show long-term persistence and potent antitumor activity

Chimeric antigen receptors (CARs) have been used to redirect the specificity of autologous T cells against leukemia and lymphoma with promising clinical results. Extending this approach to allogeneic T cells is problematic as they carry a significant risk of graft-versus-host disease (GVHD). Natural killer (NK) cells are highly cytotoxic effectors, killing their targets in a non-antigen-specific manner without causing GVHD. Cord blood (CB) offers an attractive, allogeneic, off-the-self source of NK cells for immunotherapy. We transduced CB-derived NK cells with a retroviral vector incorporating the genes for CAR-CD19, IL-15 and inducible caspase-9-based suicide gene (iC9), and demonstrated efficient killing of CD19-expressing cell lines and primary leukemia cells in vitro, with marked prolongation of survival in a xenograft Raji lymphoma murine model. Interleukin-15 (IL-15) production by the transduced CB-NK cells critically improved their function. Moreover, iC9/CAR.19/IL-15 CB-NK cells were readily eliminated upon pharmacologic activation of the iC9 suicide gene. In conclusion, we have developed a novel approach to immunotherapy using engineered CB-derived NK cells, which are easy to produce, exhibit striking efficacy and incorporate safety measures to limit toxicity. This approach should greatly improve the logistics of delivering this therapy to large numbers of patients, a major limitation to current CAR-T-cell therapies