Progress 05/01/23 to 04/30/24
Outputs
Target Audience:Graduate and undergraduate students of WVSU Biomedical Researchers,Geneticists and Drosophila Researchers Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Curcumin, derived from Curcuma longa and widely used as a dietary spice, has attracted attention for its therapeutic potential, including antioxidant, anti-inflammatory, and antimicrobial properties. Despite its established benefits, understanding the precise mechanisms of curcumin's effects on consumers remains a challenge. To address this gap, our research employed the genetic model Drosophila melanogaster and utilized transcriptomics and metabolomics tools. Our investigation identified significant alterations in 1043 genes and 73 metabolites upon curcumin supplementation, revealing genetic modulation in pathways related to antioxidants, carbohydrates, lipids, gustatory perception, and reproductive processes. Metabolites implicated in carbohydrate metabolism, amino acid biosynthesis, and biomarkers associated with neurodegenerative diseases were also identified. This study highlighted correlations between the curcumin diet, enhanced antioxidant mechanisms, amino acid metabolism, and specific physiological effects. Moreover, our project has significantly contributed to training and professional development by organizing workshops and virtual RNA sequence analysis sessions. These initiatives, integrated into the curricula of Biotechniques (BT 571) and Molecular Genetics (BT 573), enriched educational experiences and equipped participants with essential skills in modern plant breeding and bioinformatics. By reaching 1890 participants from various institutions, including Delaware State University, Florida A&M University, and Fort Valley State University, we fostered collaboration and knowledge exchange within the agricultural genetics research community, advancing the application of modern genomics tools in scientific inquiry. How have the results been disseminated to communities of interest? The results of our research on curcumin and its effects, conducted using Drosophila melanogaster as a genetic model, have been disseminated to communities of interest through several key avenues: Publications: Findings have been disseminated through peer-reviewed scientific journals specializing in genetics, molecular biology, and pharmacology. These publications detail our methodologies, results, and implications for understanding curcumin's impact on gene expression, metabolite profiles, and physiological outcomes. Conferences and Symposia: Presentations and posters have been delivered at national and international scientific conferences focused on genetics, bioinformatics, and health sciences. These platforms allow for direct engagement with researchers, practitioners, and stakeholders interested in curcumin's therapeutic potential. Workshops and Training Sessions: Our project has organized workshops and virtual sessions tailored for graduate students, undergraduate students, postdoctoral associates, and faculty. These sessions cover bioinformatics pipelines, transcriptomics, and genomic analysis techniques, fostering knowledge dissemination and skill development among future researchers. Collaborative Networks: Collaborations with other academic institutions, research centers, and industry partners have facilitated sharing results and insights. Through these networks, we exchange knowledge, validate findings, and explore further applications of curcumin in health and disease. By leveraging these dissemination strategies, we aim to maximize the impact of our research on curcumin and contribute to advancing scientific knowledge and applications in biomedical research and health sciences. What do you plan to do during the next reporting period to accomplish the goals?During the upcoming reporting period, our project at WVSU will focus on integrating microbiome and big-data analysis skills into high school and undergraduate education through a comprehensive approach. We will conduct rigorous Drosophila genomic research using wild-derived DGRP lines to investigate the effects of phytochemical diets, including capsaicin, curcumin, and carotenoids, on genetic and metabolic responses. This research will involve Genome-Wide Association Studies (GWAS) to identify key genes associated with body weight, triglyceride levels, and glucose metabolism. Simultaneously, we will launch a Genomics Summer Institute, where students will learn Drosophila rearing techniques, conduct feeding trials, assay gut microbiome composition, and analyze bioinformatics data. This hands-on training will empower students to explore the influence of diet on microbiome diversity and its health implications. Additionally, our Community-Based After School Club will engage students in ongoing microbiome investigations, fostering skills in experimental design, data interpretation, and scientific communication. These initiatives aim to advance scientific knowledge and cultivate a new generation of researchers equipped with essential skills in genomics and microbiome research.?
Impacts
What was accomplished under these goals?
Our integrated research project is not just about exploring the genetic underpinnings of body weight reduction in Drosophila through phytochemicals and investigating the responses of the Drosophila gut microbiome to diets enriched with these phytochemicals. It's also about investing in the future of science. We are committed to advancing scientific understanding through in-depth genomic research and genome-wide association studies (GWAS) utilizing phytochemicals from our pepper collections, as well as training WVSU students in cutting-edge techniques. These students are gaining hands-on experience in Drosophila rearing, conducting feeding trials, analyzing gut microbiome data, and utilizing bioinformatics tools. Our project also includes initiatives like the Metagenomics Summer Institute for Undergraduate/High School students and Community-Based After School Club Experiences, which aim to engage and educate the broader community in the applications of metagenomics and bioinformatics in health and environmental sciences. This integrated approach, which mirrors recent studies on curcumin in Drosophila, is a significant step towards bridging gaps in our understanding of the health impacts of phytochemicals, and it's a testament to our optimism about the future of the field.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2024
Citation:
Belcher S, Flores-Iga G, Natarajan P, Crummett G, Talavera-Caro A, Gracia-Rodriguez C, Lopez-Ortiz C, Das A, Adjeroh DA, Nimmakayala P, et al. Dietary Curcumin Intake and Its Effects on the Transcriptome and Metabolome of Drosophila melanogaster. International Journal of Molecular Sciences. 2024; 25(12):6559. https://doi.org/10.3390/ijms25126559
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
Belcher, S., Flores-Iga, G., Natarajan, P., Crummett, G., Talavera-Caro, A., Gracia-Rodriguez, C., Lopez-Ortiz, C., Das, A., Adjeroh, D.A., Nimmakayala, P., Balagurusamy, N., and Reddy, U.K. Curcumin Intake and Its Effects on the Transcriptome and Metabolome of Drosophila melanogaster. 12th Annual American Council For Medicinally Active Plants Conference (October 18-20, 2023).
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Progress 05/01/22 to 04/30/23
Outputs
Target Audience:Graduate and undergraduate students of WVSU Biomedical Researchers, Pepper Geneticists and Drosophila Researchers Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Graduate student and research associate working in this project attended the Drosophila genetics 2023 conference and presented their work as a poster presentation. All the project personnel are well trained in handling the flies and conducting feeding experiments, dissecting the guts andtranscriptome analysis. They will be attending the future conferences to present their work How have the results been disseminated to communities of interest?Research papers, conference presentations and classroom teaching What do you plan to do during the next reporting period to accomplish the goals?We will conduct experiments to explore the changes in gut microbiota of D. melanogaster reared on diets supplemented with phytochemicals derived from pepper fruits. Three classical wild-type strains of D. melanogaster including Oregon-RC (5), Canton-S (64349), and Berlin-K (8522) will be used for metagenomic analysis. Populations of all three genotypes will be placed on different diets: control, and phytochemical- (i.e., capsaicin, dihydrocapsaicin, capsiate, carotenoids, flavonoids,) containing diets. The phytochemical-containing diets will consist of autoclave-sterilized standard cornmeal medium (Nutri-fly Bloomington formulation) solidified with agar and supplemented with 0.4% propionic acid (v/v) and 0.3% Tegosept (w/v) as preservatives. The control diet will include this formulation except for phytochemical supplementation. All experiments and culturing will be performed under controlled conditions at 25 °C on a 12 h light/dark photoperiod. Experiments will be initiated by placing 10 males and 10 females of each genotype onto vials containing the different diets. Adults will be allowed to lay eggs for 72 h before being removed. The larvae will be fed and at the adult stage, these flies will be selected for body-weight measurement, triglyceride, and glucose level analysis. Each of the control and phytochemical-containing lines will be maintained in three independent replications of vial culture. Drosophila guts will be dissected by using a modified protocol described by Garcia-Lozano et al., (2019) and DNA will be extracted for sequencing. For amplification of the 16S rDNA gene, bacterial universal primers 341F (5'-CCTAYGGGRBGCASCAG-3') and 806R (5'-GGACTACNNGGGTATCTAAT-3') will be used to amplify the V3-V4 variable regions. PCR will involve a modified PCR protocol from initial denaturation at 94°C for 3 min followed by 45 cycles of 94°C for 45 sec, 50°C for 60 sec, and 72°C for 90 sec, with a final extension at 72°C for 10 min . Products will be checked by 1% gel electrophoresis and purified by using the Qiagen PCR Purification kit. Sequencing and data analysis will be done using the standardized procedures in our laboratory.
Impacts
What was accomplished under these goals?
As per the objectives, we performed the Genome-Wide Transcriptome and Metabolome analysesin Drosophila melanogaster using pepper and curcumin enriched diets. Populations of the Berlin-K (8522) Drosophila genotype were placed on control and habanero-containing diets. The control diet consisted of autoclave-sterilized standard cornmeal medium solidified with agar and supplemented with 0.5% propionic acid (v/v) and 1.5% Tegosept (w/v) as preservatives. The habanero-pepper diet included this formulation and also 7.5% (w/v) ground and dried habanero pepper. Experiments were initiated by placing 10 male and 10 female flies into vials containing the different diets. Adults were allowed to lay eggs for 96 h before being removed. The larvae were fed, and once the adult stage was achieved, these flies were selected for body weight, triglycerides (TG), and glucose measurements; total RNA extraction; and metabolome analysis. Each of the control and pepper diet-reared lines were maintained in three independent replications. Adult flies were placed into Eppendorf tubes in batches per each sex and stored at -80 degree Centigrade.The amount of food eaten by single flies was measured using the Capillary Feeder(CAFE) assay. We performed transcriptomic and metabolomic analyses of adult flies reared on a habanero pepper diet. We found 539 genes/59 metabolites that were differentially expressed/accumulated in flies fed a pepper versus control diet. Transcriptome results indicated that olfactory sensitivity and behavioral responses to the pepper diet were mediated by olfactory and nutrient-related genes including gustatory receptors (Gr63a, Gr66a, and Gr89a), odorant receptors (Or23a, Or59a, Or82a, and Orco), and odorant-binding proteins (Obp28a, Obp83a, Obp83b, Obp93a, and Obp99a). Metabolome analysis revealed that campesterol, sitosterol, and sucrose were highly upregulated and azelaic acid, ethyl phosphoric acid, and citric acid were the major metabolites downregulated in response to the habanero pepper diet. Further investigation by integration analysis between transcriptome and metabolome data at gene pathway levels revealed six unique enriched pathways, including phenylalanine metabolism; insect hormone biosynthesis; pyrimidine metabolism; glyoxylate, and dicarboxylate metabolism; glycine, serine, threonine metabolism; and glycerolipid metabolism. Effect of curcuminoid-enriched diet on the Diversity and Composition of gut microbiome in Drosophila melanogaster Microbial communities found in the gut of animals play an important role in the health of the animal through the breakdown of food for nutrient and energy extraction, production of essential vitamins, and protection against pathogen colonization. Microbiome composition and stability can differ depending on intrinsic factors of the host, including age, sex, genotype, and exogenous factors such as habitat and diet. One of the greatest impacts on the gastrointestinal microbiome is diet because the host and microbiome share the same food source. Curcumin is one of the major isolated curcuminoids derived from the rhizome of Curcuma longa (turmeric), a member of the ginger family (Zingiberaceae). It has been reported that curcumin can help in the management of oxidative and inflammatory conditions, metabolic syndrome (i.e., heart disease, stroke, type 2 diabetes), arthritis, anxiety, and hyperlipidemia. Diets enriched with curcuminoids might cause shifts in the microbiome. Thus, understanding how these interactions occur can reveal potential health implications associated with such changes. We conducted an experiment with adult flies, to explore the gut microbiome of differentDrosophilagenetic backgrounds and the effects of dietary curcuminoid treatments on its composition and structure. We analyzed the gut microbiomes of two Drosophila melanogastergenetic backgrounds (Oregon-RC and Berlin-K) reared on control and curcuminoid-containing diets (curcumin and turmeric). Results of 16S rRNA gene sequencing revealed that the dominant phylum observed was Firmicutes, with varying proportions, ranging from 80.28% in Oregon adult flies on a turmeric diet to a remarkable 99.47% in Oregon larva flies under the same dietary condition. Specifically, the Lactobacillaceae family within Firmicutes stood out prominently. In addition, the Proteobacteria phylum was detected across samples, spanning from 0.173% to 11.16%, while Actinobacteria appeared in the range of 0% to 1.94%. Interestingly, when comparing the two fly genotypes, Berlin-K and Oregon-RC, Proteobacteria exhibited higher abundance in the adult stage, whereas Actinobacteria were more prevalent during the larval stage. This study revealed thatcurcumin-containing diets can shift the Drosophilagut microbiome, but the response varied according to the genotype and supplemented diet.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2023
Citation:
Lopez-Ortiz C, Gracia-Rodriguez C, Belcher S, Flores-Iga G, Das A, Nimmakayala P, Balagurusamy N, Reddy UK. Drosophila melanogaster as a Translational Model System to Explore the Impact of Phytochemicals on Human Health. International Journal of Molecular Sciences. 2023; 24(17):13365. https://doi.org/10.3390/ijms241713365
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
Lopez-Ortiz, C., Edwards, M., Natarajan, P., Pacheco-Valenciana, A., Nimmakayala, P., Adjeroh, D. A., ... & Reddy, U. K. (2022). Peppers in Diet: Genome-Wide Transcriptome and Metabolome Changes in Drosophila melanogaster. International Journal of Molecular Sciences, 23(17), 9924.
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