Breast-cancer-secreted miR-122 reprograms glucose metabolism in premetastatic niche to promote metastasis

Reprogrammed glucose metabolism as a result of increased glycolysis and glucose uptake is a hallmark of cancer. Here we show that cancer cells can suppress glucose uptake by non-tumour cells in the pre-metastatic niche, by secreting vesicles that carry high levels of the miR-122 microRNA. High miR-122 levels in the circulation have been associated with metastasis in breast cancer patients and we show that cancer-cell-secreted miR-122 facilitates metastasis by increasing nutrient availability in the pre-metastatic niche. Mechanistically cancer-cell-derived miR-122 suppresses glucose uptake by niche cells in vitro and in vivo by downregulating the glycolytic enzyme pyruvate kinase (PKM). In vivo inhibition of miR-122 restores glucose uptake in distant organs, including brain and lungs, and decreases the incidence of metastasis. These results demonstrate that by modifying glucose utilization by recipient pre-metastatic niche cells, cancer-derived extracellular miR-122 is able to reprogram systemic energy metabolism to facilitate disease progression

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

Attempted Cloning of a Wnt Gene from Botrylloides violaceus

Botrylloides violaceus is a colonial ascidian with the ability to undergo sexual and asexual reproduction as well as regeneration. The canonical pathway starts with the extracellular protein Wnt and ends with β-catenin, a transcription factor, which also functions in cell adhesion. The Wnt signaling pathway is involved in embryogenesis and regeneration in a variety of other species. In our studies we attempt to isolate and sequence both a Wnt gene and from Botrylloides via degenerate primer design and PCR. Using bioinformatic methods we aligned sequences from other organisms, as the Botrylloides genome has not yet been sequenced. Using mouse, Ciona, sea urchin, and anemone DNA we were able to design primers using areas of conservation in both genes. We have been optimizing DNA and RNA isolation procedures from Botrylloides and are now identifying optimal annealing temperatures for our primers. Obtaining sequence data from fragments of these genes will allow us to compare them to sequences from other species and provide us with information about evolutionary conservation

Cold Tolerance Mechanisms in Mungbean (<i>Vigna radiata</i> L.) Genotypes during Germination

Mungbean or greengram (Vigna radiata) is an important legume crop well known for its high protein with nitrogen-fixing abilities. However, the severe yield loss in mungbean occurs due to susceptibility to low temperatures at all stages of plant growth including germination and is a serious concern for its cultivation and productivity. To select cold-tolerant genotypes, a germination-based screening at 10 °C was performed in a total of 204 germplasms. The study showed that cold stress of the initial 8-days during seedling establishment imposed a negative impact throughout the life of mungbean genotypes, which were reflected in the vegetative and reproductive phase (plant height, days to 50% flowering and pods/plant, seeds/pod, yield/plant, and 100-seed weight). The biplot analysis showed that parameters such as germination rate index, Timson’s index, mean germination time, and coefficient of the velocity of germination are the key influential germination parameters for identifying cold tolerance in the seedling stage. Identified cold-tolerant genotype (PAU911) retained higher rootlet number, leaf area, and increased chlorophyll, carotenoid, and malondialdehyde (MDA) content at 10 °C. Based on the confocal microscopic study, it is noticed that the stomatal density, open pore percentage, and trichome density were significant differences in seedlings exposed to cold stress as compared to non-stress. On the basis of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) analysis, it is observed that a new protein identified as TETRATRICOPEPTIDE-REPEAT THIOREDOXIN-LIKE1 (TTL1) (UNIPROT Identifier: LOC106762419) which highly correlated with the cold stress response of in the cold-tolerant genotype. Our study identifies a noble member, TTL1, whose expression has a positive role in cold tolerance response at the protein level in V. radiata. This study will help breeding programs with regard to the sustainable growth of mungbean

Associations between Maternal Education and Child Nutrition and Oral Health in an Indigenous Population in Ecuador

The global nutrition transition has increased the prevalence of childhood dental caries. Greater understanding is needed of the impact of social determinants&mdash;including maternal education&mdash;on child oral health. This is a cross-sectional analysis of a convenience sample of families of 458 indigenous Ecuadorian children aged 6 months through 6 years from 2011&ndash;2013. Data was collected by mother interviews and child dental and anthropometric examinations. Multivariate logistic and Zero-Inflated-Poisson regression analyses assessed associations between years of maternal education and maternal-child oral health practices and child oral health outcomes. Each additional year of maternal education was significantly (p &lt; 0.05) associated with some healthier practices including greater likelihood of mothers and children drinking milk daily (OR 1.20; 95% CI 1.08, 1.34); and less healthy practices including greater likelihood of bottle-feeding children with sugary liquids (OR 1.14; 95% CI 1.06, 1.22) and to older age, giving children sweets daily, calming children with a bottle or sweets, and less likelihood of helping brush their children&rsquo;s teeth (OR 0.93; 95% CI 0.88, 0.98). Each year of maternal education had a small but statistically non-significant influence on increasing the odds of children being among those who are cavity-free (OR 1.03; 95% CI 0.92, 1.16). Interventions to improve health outcomes should focus not just on maternal education but also address social and commercial determinants of health through nutrition and oral health education, as well as policies to reduce sugar and ensure universal access to oral health care

Optimizing the transduction efficiency of capsid-modified AAV6 serotype vectors in primary human hematopoietic stem cells in vitro and in a xenograft mouse model in vivo

Background aimsAlthough recombinant adeno-associated virus serotype 2 (AAV2) vectors have gained attention because of their safety and efficacy in numerous phase I/II clinical trials, their transduction efficiency in hematopoietic stem cells (HSCs) has been reported to be low. Only a few additional AAV serotype vectors have been evaluated, and comparative analyses of their transduction efficiency in HSCs from different species have not been performed.MethodsWe evaluated the transduction efficiency of all available AAV serotype vectors (AAV1 through AAV10) in primary mouse, cynomolgus monkey and human HSCs. The transduction efficiency of the optimized AAV vectors was also evaluated in human HSCs in a murine xenograft model in vivo.ResultsWe observed that although there are only six amino acid differences between AAV1 and AAV6, AAV1, but not AAV6, transduced mouse HSCs well, whereas AAV6, but not AAV1, transduced human HSCs well. None of the 10 serotypes transduced cynomolgus monkey HSCs in vitro. We also evaluated the transduction efficiency of AAV6 vectors containing mutations in surface-exposed tyrosine residues. We observed that tyrosine (Y) to phenylalanine (F) point mutations in residues 445, 705 and 731 led to a significant increase in transgene expression in human HSCs in vitro and in a mouse xenograft model in vivo.ConclusionsThese studies suggest that the tyrosine-mutant AAV6 serotype vectors are the most promising vectors for transducing human HSCs and that it is possible to increase further the transduction efficiency of these vectors for their potential use in HSC-based gene therapy in humans

Breast cancer-secreted miR-122 reprograms glucose metabolism in pre-metastatic niche to promote metastasis

Reprogrammed glucose metabolism as a result of increased glycolysis and glucose uptake is a hallmark of cancer. Here we show that cancer cells can suppress glucose uptake by non-tumour cells in the pre-metastatic niche, by secreting vesicles that carry high levels of the miR-122 microRNA. High miR-122 levels in the circulation have been associated with metastasis in breast cancer patients and we show that cancer-cell-secreted miR-122 facilitates metastasis by increasing nutrient availability in the pre-metastatic niche. Mechanistically cancer-cell-derived miR-122 suppresses glucose uptake by niche cells in vitro and in vivo by downregulating the glycolytic enzyme pyruvate kinase (PKM). In vivo inhibition of miR-122 restores glucose uptake in distant organs, including brain and lungs, and decreases the incidence of metastasis. These results demonstrate that by modifying glucose utilization by recipient pre-metastatic niche cells, cancer-derived extracellular miR-122 is able to reprogram systemic energy metabolism to facilitate disease progression