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Reddy, S.

Publications and source records attributed to Reddy, S..

41 records · Page 3Linked to original sources

Immune responses in pancreatic cancer may be restricted byprevalence of activated regulatory T-cells, dysfunctionalCD8+ T-cells, and senescent T-cells

Pancreatic cancer has the worst prognosis of any human malignancy and leukocyte infiltration is a major prognostic marker of the disease. As current immunotherapies confer negligible survival benefits, there is a need to better characterise leukocytes in pancreatic cancer to identify better therapeutic strategies. In this study, we analysed 32 human pancreatic cancer patients from two independent cohorts. A multi-parameter mass-cytometry analysis was performed on 32,000 T-cells from eight patients. Single-cell RNA sequencing dataset analysis was performed on a cohort of 24 patients. Multiplex immunohistochemistry imaging and spatial analysis were performed to map immune infiltration into the tumour microenvironment. Regulatory T-cell populations demonstrated highly immunosuppressive states with high TIGIT, ICOS and CD39 expression. CD8+ T-cells were found to be either in senescence or an exhausted state. The exhausted CD8 T-cells had low PD-1 expression but high TIGIT and CD39 expression. These findings were corroborated in an independent pancreatic cancer single-cell RNA dataset from additional 24 patients. These data suggest that T-cells are major players in the suppressive microenvironment of pancreatic cancer. Our work identifies novel therapeutic targets that should form the basis for rational design of a new generation of clinical trials in pancreatic ductal adenocarcinoma.

immunology↗

Human intestinal tissue-resident memory CD8+ T cells comprise transcriptionally and functionally distinct subsets

Tissue-resident memory T (TRM) cells provide key adaptive immune responses in infection, cancer, and autoimmunity. However transcriptional heterogeneity of human intestinal TRM cells remains undefined, and definitive markers of CD103-TRM cells are lacking. Here, we investigated transcriptional and functional heterogeneity of human TRM cells through the study of donor-derived intestinal TRM cells from intestinal transplant recipients. Single-cell transcriptional profiling identified four conventional TRM populations, with two distinct transcriptional states of CD8+ TRM cells, delineated by ITGAE and ITGB2 expression. We defined a transcriptional signature discriminating the two CD8+ populations, including differential expression of key residency-associated genes and cytotoxic molecules. Flow cytometry of recipient-derived cells infiltrating the graft and intestinal lymphocytes from healthy gut confirmed the two CD8+ TRM phenotypes, with {beta}2-integrin acting as a CD103-CD8+ TRM marker. CD103+ CD8+ TRM cells produced IL-2, and demonstrated greater polyfunctional cytokine production, while {beta}2-integrin+ CD69+ CD103-TRM cells had higher granzyme expression. Phenotypic and functional analysis of intestinal CD4+ T cells identified many parallels, including a distinct {beta}2-integrin+ population. Together, these results describe the transcriptional, phenotypic, and functional heterogeneity of human intestinal TRM cells, and suggest a role for {beta}2-integrin in TRM development. SummaryHeterogeneity within human tissue-resident memory T (TRM) cells is poorly understood. We show that transcriptionally, phenotypically, and functionally distinct CD4+ and CD8+ TRM subsets exist in the human intestine, and that {beta}2-integrin expression identifies a distinct population of CD8+ TRM cells.

immunology↗

Diversification of a cryptic radiation, a closer look at Madagascar’s recently recognized bird family

Despite its status as a global biodiversity hotspot there is still much to be discovered about the birds of Madagascar, including a full accounting of species-level diversity and the avifaunas origins. The Bernieridae is a Malagasy endemic family that went unrecognized by science for decades and unnamed until 2010. This cryptic family has long represented a missing piece of the puzzle of the avian tree of life. We present the first comprehensive phylogeny of Bernieridae in order to examine its diversification history on Madagascar and its place within Passeriformes. In light of recent discoveries of cryptic species-level diversity in Madagascars vertebrate fauna, we used broad geographic sampling and integrative taxonomic methods to investigate the potential for cryptic lineages within every known species of the Bernieridae. Our approach combines phylogenomics using [~]4500 loci of ultraconserved elements (UCEs), genetic clustering of thousands of single nucleotide polymorphisms (SNPs), and statistical analysis of morphological variation. These methods yielded the discovery of two unrecognized species in the previously monotypic genus Bernieria, along with new insights into patterns of fine-scale endemism in Madagascars humid forests. Our phylogenomic analyses provide conclusive support for Donacobiidae and Bernieridae as sister families, a biogeographically intriguing result given that the former is restricted to the Neotropics. We found a significant decline in the rate of speciation over time on Madagascar, consistent with a model of adaptive radiation. Bernieridae therefore joins the Vangidae as a second avian adaptive radiation on the island of Madagascar. These insights into the evolution of Bernieridae represent a step forward in understanding the origins and diversity of Madagascars endemic avifauna.

evolutionary biology↗

Phosphoprotein-based Biomarkers as Predictors for Cancer Therapy

Disparities in cancer patient responses have prompted widespread searches to identify differences in sensitive vs. non-sensitive populations and form the basis of personalized medicine. This customized approach is dependent upon the development of pathway-specific therapeutics in conjunction with biomarkers that predict patient responses. Here, we show that Cdk5 drives growth in subgroups of human patients with multiple types of neuroendocrine neoplasms. Phosphoproteomics and high throughput screening identified novel phosphorylation sites downstream of Cdk5. These phosphorylation events serve as biomarkers of Cdk5 activity and effectively pinpoint Cdk5-driven tumors. Toward achieving targeted therapy, we demonstrate that mouse models of neuroendocrine cancer are responsive to selective Cdk5 inhibitors and biomimetic nanoparticles are effective vehicles for enhanced tumor targeting and reduction of drug toxicity. Finally, we show that biomarkers of Cdk5-dependent tumors effectively predict response to anti-Cdk5 therapy in patient-derived xenografts. Thus, a phosphoprotein-based diagnostic assay combined with Cdk5-targeted therapy is a rational treatment approach for neuroendocrine malignancies.

cancer biology↗

Preparation of Artesunate mPEG-PLGA Nanoparticles and Its Application to K562 Apoptosis of cells

The preparation process of artesunate-loaded polyethylene glycol monomethyl ether-polylactic acid-glycolic acid affinity block copolymer (mPEG-PLGA) nanoparticles and its growth inhibition on human leukemia K562 cells were investigated. METHODS: Artesunate mPEG-PLGA nanoparticles (Art-Nps) were prepared by modified self-emulsification method. The morphology of nanoparticles was characterized by scanning electron microscopy. The particle size distribution and zeta potential were measured by laser scattering particle size analyzer. The drug loading, encapsulation efficiency and in vitro release of Art-Nps were determined by chromatography. The proliferation and apoptosis of human leukemia K562 cells were observed by MTT assay and Hoechst staining. RESULTS: Art-Nps is a spherical solid particle with smooth surface, average particle size (156.70+/-1.01) nm, zeta potential of -(26.23+/-1.86) mV, average drug loading (14.51+/-0.20)%, average package. The sealing rate was (86.51+/-0.50)%, and the in vitro release law accorded with the Higuchi equation: Q=4.11t 1/2+27.05, R2=0.98. MTT assay showed that Art-Nps inhibited the proliferation of K562 cells in a time-dose-dependent manner, and the inhibition rate exceeded the artesunate-treated group after 72h, and sustained release. The number of cells was observed after cultured with different concentrations of Art-Nps for 48h. Significantly reduced, cell size is different, irregular shape, high magnification can be seen in the nucleus pyknosis, agglutination, and apoptotic bodies, and increased apoptotic bodies with increasing concentration.

bioengineering↗