Biology subjects

Martins, A. J.

Publications and source records attributed to Martins, A. J..

10 recordsLinked to original sources

bioRxiv2025

Cytochrome P450 inhibition impedes pyrethroid effects on insects through Nav channel regulation

Insecticides used in various formulations are inevitable in agricultural pest control and prevention of vector-borne diseases. Piperonyl butoxide (PBO) is a synergist widely used to enhance the effectiveness of insecticides, notably pyrethroids, by inhibiting the detoxifying cytochrome P450 enzymes, thus reducing the capacity of insects to metabolize and resist insecticides. Recent studies, however, reveal an unexpectedly weak restoration of pyrethroid efficacy by PBO, but the underlying mechanism is unknown. Here, we demonstrate that the PBO-induced inhibition of cytochrome P450 impedes the effect of deltamethrin by mainly affecting its interaction with the inactivated state of voltage-gated sodium channels (Nav). We describe a new octopamine-dependent regulatory mechanism involving Gs, PKA, DARPP-32, and PP1-2A, which affect the cytochrome P450 conformation, thus limiting the effect of PBO and modulating the Nav gating. As a result, deltamethrin cannot reach its final binding site in the fenestration to exert its full effect. We confirmed in vivo that under chemical stressor, the level of octopamine is elevated, which decreases the deltamethrin efficacy. Our findings reveal a novel adaptation mechanism that increases insect survival by reducing insecticide efficacy, thus emphasizing the necessity of developing more effective formulations and technologies for pest and vector control.

pharmacology and toxicology↗

bioRxiv2024

Single-nucleus RNA sequencing of human periventricular white matter in vascular dementia

Vascular dementia (VaD) refers to a variety of dementias driven by cerebrovascular disease and is the second leading cause of dementia globally. VaD may be caused by ischemic strokes, intracerebral hemorrhage, and/or cerebral small vessel disease, commonly identified as white matter hyperintensities on MRI. The mechanisms underlying these white matter lesions in the periventricular brain are poorly understood. In this study we perform an extensive transcriptomic analysis on human postmortem periventricular white matter lesions in patients with VaD with the goal of identifying molecular pathways in the disease. We find increased cellular stress responses in astrocytes, oligodendrocytes, and oligodendrocyte precursor cells as well as transcriptional and translational repression in microglia in our dataset. We show that several genes identified by GWAS as being associated with white matter disease are differentially expressed in cells in VaD. Finally, we compare our dataset to an independent snRNAseq dataset of PVWM in VaD and a scRNAseq dataset on human iPSC-derived microglia exposed to oxygen glucose deprivation (OGD). We identify the increase of the heat shock protein response as a conserved feature of VaD across celltypes and show that this increase is not linked to OGD exposure. Overall, our study is the first to show that increased heat shock protein responses are a common feature of lesioned PVWM in VaD and may represent a potential therapeutic target.

neuroscience↗

bioRxiv2024

The Multiple Roles of Gamma Interferon in Intraepithelial T Cell-Villous Enterocyte Interactions in Active Celiac Disease

We compared duodenal biopsies showing active celiac disease (CeD) to normal controls using single cell RNA sequencing, cyclic immunofluorescence, RNAScope and proximity ligation assays. There is increased infiltration of villous but not crypt epithelium T cells bearing either {beta} or {gamma}{delta} T cell receptors (TCRs) in CeD. Both T cell subsets are activated cytotoxic T lymphocytes (CTLs) and surprisingly are the predominant mucosal source of IFN{gamma}. In response to this IFN{gamma},villous but not crypt enterocytes show an IFN{gamma} signature, including nuclear phospho-STAT1 protein, class II HLA molecules and IFN{gamma}-inducible chemokines known to recruit CTLs (e.g., CCL3, CCL4, CXCL10, and CXCL11) and receptors for these chemokines are expressed on the infiltrating CTLs. Villous enterocytes also display increased HLA-E and HLA-B mRNAs and proteins. Bioinformatic analyses (NICHES) and proximity ligation assays show frequent binding of both {beta} and {gamma}{delta} TCRs with enterocyte HLA-E or HLA-B, but not HLA-DR. In contrast, NKG2C, proposed as an alternative trigger of CTL activation, is infrequently-expressed and shows few interactions with HLA-E. Our data suggest that activated intraepithelial CTLs produce IFN{gamma} which recruits additional CTLs and increases antigen-dependent killing of villous epithelium using either conventional or HLA-E antigen presentation. SignifanceAnalyses of diagnostic biopsies from celiac disease patients reveals critical roles for IFN{gamma} actions on enterocytes and a surprising role of HLA-E in antigen presntation. Activated cytotoxic T lymphocytes (CTLs) that are the primary source of mucosal IFN{gamma}, leading to IFN{gamma} signaling in villous enterocytes, including expression of phospho-STAT1 protein, class II HLA molecules, and chemokines that attract CTLs. Bioinformatic ligand receoptor analyses and proximity ligation assays showed frequent interactions between intraepithelial CTLs and enterocyte HLA-E or HLA-B, indicating that IFN{gamma} production by the CTLs likely promotes further CTL recruitment and antigen-dependent killing of the villous epithelium.

immunology↗

bioRxiv2024

Comprehensive Molecular Epidemiology of Influenza Viruses in Brazil: Insights from a Nationwide Analysis

Influenza A and B viruses pose significant global health threats, with substantial impacts on morbidity and mortality. Understanding their molecular epidemiology in Brazil, a key hub for the circulation and dissemination of these viruses in South America, remains limited. This study, part of the Center for Viral Surveillance and Serological Assessment (CeVIVAS) project, addresses this by analyzing data and samples from all Brazilian macroregions, along with publicly available sequences from 2021-2023. Phylogenetic analysis of the Hemagglutinin (HA) segment of Influenza A/H1N1pdm09, A/H3N2, and Influenza B/Victoria-lineage revealed the predominance of A/H3N2 2a.3 strain in 2021 and early 2022. This was succeeded by A/H3N2 2b until October 2022, after which A/H1N1pdm09 5a.2a and 5a.2a.1 lineages became prevalent, maintaining this status throughout 2023. B/Victoria circulated at low levels between December 2021 and September 2022, becoming co-prevalent with A/H1N1pdm09 5a.2a and 5a.2a.1 lineages. Comparing the vaccine strain A/Darwin/9/2021 with circulating A/H3N2 viruses from 2021-2023 revealed shared mutations to aspartic acid at residues 186 and 225, altering the RBD domains charge. For A/H1N1pdm09, the 2022 consensus of 5a.2a.1 and the vaccine strain A/Victoria/2570/2019 had 14 amino acid substitutions. Key residues such as H180, D187, K219, R223, E224, and T133 are involved in hydrogen interactions with sialic acids, while N130, K142, and D222 may influence distance interactions based on docking analyses. Distinct Influenza A lineage frequency patterns across Brazils macroregions underscore regional variations in virus circulation. This study characterizes the dynamics of Influenza A and B viruses in Brazil, offering valuable insights into their circulation patterns. These findings have significant public health implications, informing strategies to mitigate transmission risks, optimize vaccination efforts, and enhance outbreak control measures. Author summaryThis study investigates the molecular epidemiology of Influenza A and B viruses in Brazil from 2021 to 2023. Utilizing data from the Center for Viral Surveillance and Serological Assessment (CeVIVAS) and public databases, we performed a comprehensive phylogenetic analysis of the Hemagglutinin segments of Influenza A/H1N1pdm09, A/H3N2, and B/Victoria-lineage viruses across all Brazilian macroregions. Key findings reveal that the A/H3N2 2a.3 strain was predominant in 2021 and early 2022, followed by A/H3N2 2b, and later by A/H1N1pdm09 5a.2a and 5a.2a.1 lineages in late 2022 and throughout 2023. The B/Victoria strain circulated at low levels initially and later co-prevailed with A/H1N1pdm09 lineages. Comparing the vaccine strain A/Darwin/9/2021 with circulating A/H3N2 viruses from 2021-2023 and A/Victoria/2570/2019 with 5a.2a.1 of A/H1N1pdm09 circulating in 2022 revealed significant mutations which could affect the interaction of the viruses with sialic acids and potentially impact vaccine efficacy. Notably, we identified a substitution pattern among the predominant Influenza subtypes and observed distinct regional variations in Influenza A lineage frequencies across Brazil. These findings are critical for optimizing vaccination strategies and provide valuable data to inform public health policy and improve health outcomes.

evolutionary biology↗

bioRxiv2023

A ticket to ride: genetic structure and kdr mutations in Aedes aegypti populations along a road crossing the Amazon Rainforest in Amapa State, Brazil

Insecticide resistance in Aedes aegypti is a major threat to the control of this mosquito species that transmits viruses that cause diseases like dengue, Zika, and chikungunya. One type of resistance is caused by alterations in the Nav gene, known as kdr mutations. In Brazil, different kdr haplotypes are present in Ae. aegypti and they may impede vector control operations based on pyrethroids. Although natural populations tend to accumulate genetic differences among isolated localities, this mosquito can actively and passively disperse, hitchhiked with human transportation. In this study, we investigated the genetic structure and kdr dispersion in Ae. aegypti populations in six localities of the Amapa State, Brazil located along a north-south transect of the Amazonian Forest. Genetic structure was assessed using 12 microsatellite loci in a fragment analysis sequencing procedure, and qPCR methods were used to detect the presence and frequency of three well known kdr mutations (V410L, V1016I and F1534C). We found a high prevalence of kdr alleles in all localities, indicating that kdr is spreading in the Amapa State. The microsatellite analyses suggested a certain level of differentiation among the mosquito populations, dividing them into two well-defined clusters, as evidenced by Bayesian and DAPC analyses. The population from Oiapoque (located in the north along the border with French Guiana) had the highest kdr frequencies and the highest genetic differentiation compared to the other localities. Our findings suggest that there is genetic structure among Ae. aegypti from the Amapa State, but with some level of passive gene flow between population clusters. The study highlights the importance of continued surveillance of Ae. aegypti populations to monitor the spread of insecticide resistance and inform on vector control strategies. Author summaryAmapa, a State in northern Brazil, is crucial as a gateway for diseases due to its border with French Guiana. One notable example was chikungunya, transmitted by the Aedes aegypti mosquito, also responsible for dengue, Zika, and yellow fever viruses. Chemical insecticides have been the primary means of control, however, resistance to these compounds is spreading among Ae. aegypti populations, compromising their effectiveness. Mutations known as "kdr" contribute to resistance to pyrethroid insecticides. Our study focused on these mutations and genetic differentiation in Ae. aegypti populations from six cities along the road nestled amidst the Amazonian Forest, linking the capital, Macapa, to Oiapoque on the Brazil-French Guiana border. The results revealed high frequencies of kdr mutations in all populations, indicating probable resistance to pyrethroids. Genetic analysis showed two distinct groups among the mosquitoes, with evidence of mosquito flow, particularly between Macapa and Oiapoque, facilitated by human transportation. Monitoring insecticide resistance and mosquito migration is essential for effective vector control strategies. Understanding these factors enables us to combat mosquito-borne diseases and safeguard public health.

evolutionary biology↗

bioRxiv2022

Multiplex high resolution melting PCR for simultaneous genotyping of pyrethroid-resistance associated mutations in Aedes aegypti. First report on kdr mutations in wild populations from Argentina

BackgroundAedes aegypti is an urban mosquito vector of Dengue and other arboviruses. During epidemic periods, pyrethroid insecticides are used for the control of adult mosquitoes; the worldwide distributed resistance to these insecticides is a cause of failures in vector control campaigns. The primary target of pyrethroids is the voltage-gated sodium channel; point mutations on this channel, called kdr mutations, are associated with pyrethroid resistance. Two kdr mutations, called V1016I and F1534C, augmented in frequency in natural populations of Ae. aegypti from the Americas in the last decade. The diagnostic of kdr polymorphisms allows an early detection of insecticide resistance spreading, which is critical for timely decisions on vector management. Given the relevance of resistance management, high-throughput methods for kdr genotyping are invaluable tools for resistance monitoring programs. These methods should also be cost-effective, to allow regional-scale surveys. Despite the extended presence of Ae. aegypti and the incidence of dengue in Argentina, the presence, abundance and distribution of kdr mutations were not reported in this country up to date. Methodology and findingsWe report a multiplex high-throughput assay based in High Resolution Melting PCR for the simultaneous genotyping of 1016 and 1534 sites in voltage-gated sodium channel gene. We used this method for the study of individual mosquito samples collected in localities which received different selection pressure with pyrethroids. Compared to other genotyping methods, multiplex High Resolution Melting was high-throughput, cost-efficient, sensitive and specific. We demonstrate for the first time the presence of kdr mutations in Argentina in regions under different selection pressure with pyrethroids. Conclusions and SignificanceWe have developed a high-throughput method for the genotyping of alleles associated with pyrethroid resistance in Ae. aegypti from the American continent. The method developed here is comparable in its sensitivity and reliability with other genotyping methods, but reduces costs and running time. It could be incorporated in control campaigns for control the presence and spreading of resistance-associated alleles. We report here for the first time the presence of kdr mutations in distant populations from Argentina, with different epidemiological situations and different history of mosquito control efforts. Authors summaryAedes aegypti is a mosquito vector of viruses such as dengue, causing millions of infections yearly worldwide. Emergence and distribution of insecticide resistance in this mosquito is a challenge for control campaigns. In this context, the implementation of resistance management strategies became a requisite for successful and sustainable mosquito management. To achieve this objective, it is important to count with early genetic markers of resistance, such as kdr mutations, which are associated with pyrethroid resistance phenotype. Kdr markers can be detected by molecular genotyping diagnostic at early stages in the emergence of resistance; this information should be considered in rational design of control campaigns. Here we present a high-throughput cost-effective method to detect kdr mutations in Ae. aegypti. Also, we demonstrate for the first time the presence of kdr mutations in Argentina.

genetics↗

bioRxiv2022

Contrasting autoimmune and treatment effects reveals baseline set points of immune toxicity following checkpoint inhibitor treatment

Immune checkpoint inhibitors (ICIs) have changed the cancer treatment landscape, but severe immune-related adverse events (irAEs) can be life-threatening or fatal and may prohibit patients from receiving further ICI treatment. While the clinical features of irAEs are well documented, molecular signatures, predictive biomarkers, and mechanisms of impending irAEs are largely unknown. In addition, the markers and mechanisms of ICI-induced antitumor immunity often overlap with those for irAEs. It is thus critical to uncover signatures associated specifically with irAEs but not with antitumor immunity. To identify circulating immune cell states associated with irAEs, we applied multimodal single cell analysis (CITE-seq) to simultaneously measure the transcriptome and surface proteins from peripheral blood mononuclear cells (PBMCs) collected before and after treatment with an anti-PD-L1 antibody (avelumab) in patients with thymic cancers (thymic epithelial tumors). All patients had an antitumor response, yet a subset developed muscle autoimmunity (myositis), a potentially life-threatening irAE. Mixed-effect modeling disentangled cell type-specific transcriptional states associated with ICI treatment responses from those of irAEs to identify temporally stable pre-treatment immune set points associated with irAEs only. These pre-treatment baseline signatures of irAE developed post-avelumab irAEs reflect correlated transcriptional states of multiple innate and adaptive immune cell populations, including elevation of metabolic genes downstream of mTOR signaling in T-cell subsets. Together these findings suggest putative pre-treatment biomarkers for irAEs following ICI therapy in thymic cancer patients and raise the prospect of therapeutically dampening autoimmunity while sparing antitumor activity in cancer patients treated with ICIs. Together, pre-treatment biomarkers and interventional therapeutics could help mitigate treatment discontinuation and improve clinical outcomes.

immunology↗

bioRxiv2021

Pre-existing chromatin accessibility and gene expression differences among naïve CD4+ T cells influence effector potential

CD4+ T cells have a remarkable potential to differentiate into diverse effector lineages following activation. Here, we probed the heterogeneity present among naive CD4+ T cells before encountering their cognate antigen to ask whether their effector potential is modulated by pre-existing transcriptional and epigenetic differences. Using single-cell RNA sequencing, we showed that key drivers of variability are genes involved in T cell receptor (TCR) signaling. Using CD5 expression as a read-out of the strength of tonic TCR interactions with self-peptide MHC, and sorting on the ends of this self-reactivity spectrum, we find that pre-existing transcriptional differences among naive CD4+ T cells impact follicular helper cell (TFH) versus non-TFH effector lineage choice. Moreover, our data implicate TCR signal strength during thymic development in establishing differences in naive CD4 T cell chromatin landscapes that ultimately shape their effector potential.

immunology↗

bioRxiv2020

Blood gene expression-based prediction of lethality after respiratory infection by influenza A virus in mice

Lethality after respiratory infection with influenza A virus (IAV) is associated with potent immune activation and lung tissue damage. In a well-controlled animal model of infection, we sought to determine if one could predict lethality using transcriptional information obtained from whole blood early after influenza virus exposure. We started with publicly available transcriptomic data from the lung, which is the primary site of the infection and pathology, to derive a multigene transcriptional signature of death reflective of innate inflammation associated with tissue damage. We refined this affected tissue signature with data from infected mouse and human blood to develop and validate a machine learning model that can robustly predict survival in mice after IAV challenge using data obtained from as little as 10 l of blood from early time points post infection. Furthermore, in genetically identical, cohoused mice infected with the same viral bolus, the same model can predict the lethality of individual animals but, intriguingly, only within a specific time window that overlapped with the early effector phase of adaptive immunity. These findings raise the possibility of predicting disease outcome in respiratory virus infections with blood transcriptional data and pave the way for translating such approaches to humans.

immunology↗

bioRxiv2020

Normalizing and denoising protein expression data from droplet-based single cell profiling

Multimodal single-cell protein and transcriptomic profiling (e.g. CITE-seq) holds promise for comprehensive dissection of cellular heterogeneity, yet protein counts measured by oligo-conjugated-antibody can have substantial noise that masks biological variations. Here we integrated experiments and computational analysis to reveal two major noise sources: protein-specific noise from unbound antibodies and cell-specific noise captured by the shared variance of isotype controls and background protein counts. We provide an open source R package (dsb) to denoise and normalize CITE-seq data based on these findings. (https://cran.r-project.org/web/packages/dsb/index.html).

bioinformatics↗