Browsing by Author "Fokar, Mohamed (TTU)"
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Item Airborne environmental DNA metabarcoding detects more diversity, with less sampling effort, than a traditional plant community survey(2021) Johnson, Mark D. (TTU); Fokar, Mohamed (TTU); Cox, Robert D. (TTU); Barnes, Matthew A. (TTU)Background: Airborne environmental DNA (eDNA) research is an emerging field that focuses on the detection of species from their genetic remnants in the air. The majority of studies into airborne eDNA of plants has until now either focused on single species detection, specifically only pollen, or human health impacts, with no previous studies surveying an entire plant community through metabarcoding. We therefore conducted an airborne eDNA metabarcoding survey and compared the results to a traditional plant community survey. Results: Over the course of a year, we conducted two traditional transect-based visual plant surveys alongside an airborne eDNA sampling campaign on a short-grass rangeland. We found that airborne eDNA detected more species than the traditional surveying method, although the types of species detected varied based on the method used. Airborne eDNA detected more grasses and forbs with less showy flowers, while the traditional method detected fewer grasses but also detected rarer forbs with large showy flowers. Additionally, we found the airborne eDNA metabarcoding survey required less sampling effort in terms of the time needed to conduct a survey and was able to detect more invasive species than the traditional method. Conclusions: Overall, we have demonstrated that airborne eDNA can act as a sensitive and efficient plant community surveying method. Airborne eDNA surveillance has the potential to revolutionize the way plant communities are monitored in general, track changes in plant communities due to climate change and disturbances, and assist with the monitoring of invasive and endangered species.Item Amino acid transporter SLC38A5 is a tumor promoter and a novel therapeutic target for pancreatic cancer(2023) Sniegowski, Tyler (TTUHSC); Rajasekaran, Devaraja (TTUHSC); Sennoune, Souad R. (TTUHSC); Sunitha, Sukumaran (TTU); Chen, Fang (TTU); Fokar, Mohamed (TTU); Kshirsagar, Sudhir (TTUHSC); Reddy, P. Hemachandra (TTUHSC); Korac, Ksenija (TTUHSC); Mahmud Syed, Mosharaf (TTUHSC); Sharker, Tanima (TTUHSC); Ganapathy, Vadivel (TTUHSC); Bhutia, Yangzom D. (TTUHSC)Pancreatic ductal adenocarcinoma (PDAC) cells have a great demand for nutrients in the form of sugars, amino acids, and lipids. Particularly, amino acids are critical for cancer growth and, as intermediates, connect glucose, lipid and nucleotide metabolism. PDAC cells meet these requirements by upregulating selective amino acid transporters. Here we show that SLC38A5 (SN2/SNAT5), a neutral amino acid transporter is highly upregulated and functional in PDAC cells. Using CRISPR/Cas9-mediated knockout of SLC38A5, we show its tumor promoting role in an in vitro cell line model as well as in a subcutaneous xenograft mouse model. Using metabolomics and RNA sequencing, we show significant reduction in many amino acid substrates of SLC38A5 as well as OXPHOS inactivation in response to SLC38A5 deletion. Experimental validation demonstrates inhibition of mTORC1, glycolysis and mitochondrial respiration in KO cells, suggesting a serious metabolic crisis associated with SLC38A5 deletion. Since many SLC38A5 substrates are activators of mTORC1 as well as TCA cycle intermediates/precursors, we speculate amino acid insufficiency as a possible link between SLC38A5 deletion and inactivation of mTORC1, glycolysis and mitochondrial respiration, and the underlying mechanism for PDAC attenuation. Overall, we show that SLC38A5 promotes PDAC, thereby identifying a novel, hitherto unknown, therapeutic target for PDAC.Item Analysis of genetic diversity and population structure of Moroccan date palm (Phoenix dactylifera L.) using SSR and DAMD molecular markers(2023) Ibrahimi, Maha; Brhadda, Najiba; Ziri, Rabea; Fokar, Mohamed (TTU); Iraqi, Driss; Gaboun, Fatima; Labhilili, Mustapha; Habach, Aicha; Meziani, Reda; Elfadile, Jamal; Abdelwahd, Rabha; Diria, GhizlaneBackground: Date palm, oasis pivot, plays a vital socio-economic part in the southern area of Morocco. However, with climate change and drought intensity and frequency increasing, the Moroccan palm grove is threatened with significant genetic degradation. Genetic characterization of this resource is key element for the development of effective conservation and management strategies in the current circumstances of climate change and various biotic and abiotic stresses. To evaluate the genetic diversity of date palm populations collected from different Moroccan oases, we used simple sequence repeats (SSR) and directed amplification of mini-satellite DNA (DAMD) markers. Our results showed that used markers could efficiently assess genetic diversity in Phoenix dactylifera L. Results: A total of 249 and 471 bands were respectively scored for SSR and DAMD, of which 100% and 92.9% were polymorphic. The polymorphic information content (PIC = 0.95), generated by the SSR primer was nearly identical to that generated by the DAMD primer (PIC = 0.98). The resolving power (Rp) was higher in DAMD than SSR (29.46 and 19.51, respectively). Analysis of the molecular variance (AMOVA) based on the combined data sets for both markers revealed a higher variance within populations (75%) than among populations (25%). Principal coordinate analysis (PCoA) and the ascendant hierarchical classification showed that the population of Zagora and Goulmima regions were the closest populations. The STRUCTURE analysis clustering of the 283 tested samples into seven clusters based on their genetic composition. Conclusion: The results drawn from this study will orient genotypes selection strategies for a successful future breeding and conservation program, particularly under climate change context.Item Chromosome-scale assembly uncovers genomic compartmentation of Fusarium oxysporum f. sp. albedinis, the causal agent of Bayoud disease in date palm(2023) Khayi, Slimane; Armitage, Andrew D.; Gaboun, Fatima; Meftah-kadmiri, Issam; Lahlali, Rachid; Fokar, Mohamed (TTU); Mentag, RachidDate palm (Phoenixdactylifera) is the most significant crop across North Africa and the Middle East. However, the crop faces a severe threat from Bayoud disease caused by the fungal pathogen Fusarium oxysporum f. sp. albedinis (FOA). FOA is a soil-borne fungus that infects the roots and vascular system of date palms, leading to widespread destruction of date palm plantations in North Africa over the last century. This is considered the most devastating pathogen of oasis agriculture in North Africa and responsible for loss of 13 million trees in Algeria and Morocco alone. In this study, we present a chromosome-scale high-quality genome assembly of the virulent isolate Foa 44, which provides valuable insights into understanding the genetic basis of Bayoud disease. The genome assembly consists of 11 chromosomes and 40 unplaced contigs, totalling 65,971,825 base pairs in size. It exhibits a GC ratio of 47.77% and a TE (transposable element) content of 17.30%. Through prediction and annotation, we identified 20,416 protein-coding genes. By combining gene and repeat densities analysis with alignment to Fusarium oxysporum f. sp. lycopersici (FOL) 4287 isolate genome sequence, we determined the core and lineage-specific compartments in Foa 44, shedding light on the genome structure of this pathogen. Furthermore, a phylogenomic analysis based on the 3,292 BUSCOs core genome revealed a distinct clade of FOA isolates within the Fusarium oxysporum species complex (FOSC). Notably, the genealogies of the five identified Secreted In Xylem (SIX) genes (1, 6, 9, 11 and 14) in FOA displayed a polyphyletic pattern, suggesting a horizontal inheritance of these effectors. These findings provide a valuable genomics toolbox for further research aimed at combatting the serious biotic constraints posed by FOA to date palm. This will pave the way for a deeper understanding of Bayoud disease and facilitate the development of effective diagnostic tools and control measures.Item Comparative Transcriptomics and Proteomics of Cancer Cell Lines Cultivated by Physiological and Commercial Media(2022) Wang, Junyao (TTU); Peng, Wenjing (TTU); Yu, Aiying (TTU); Fokar, Mohamed (TTU); Mechref, Yehia (TTU)Aiming to reduce the gap between in vitro and in vivo environment, a complex culture medium, Plasmax, was introduced recently, which includes nutrients and metabolites with concentrations normally found in human plasma. Herein, to study the influence of this medium on cellular behaviors, we utilized Plasmax to cultivate two cancer cell lines, including one breast cancer cell line, MDA-MB-231BR, and one brain cancer cell line, CRL-1620. Cancer cells were harvested and prepared for transcriptomics and proteomics analyses to assess the discrepancies caused by the different nutritional environments of Plasmax and two commercial media: DMEM, and EMEM. Total RNAs of cells were extracted using mammalian total RNA extract kits and analyzed by next-generation RNA sequencing; proteomics analyses were performed using LC-MS/MS. Gene oncology and pathway analysis were employed to study the affected functions. The cellular invasion and cell death were inhibited in MDA-MB-231BR cell line when cultured in Plasmax compared to DMEM and EMEM, whereas the invasion, migration and protein synthesis of CRL-1620 cell line were activated in Plasmax in relative to both commercial media. The expression changes of some proteins were more significant compared to their corresponding transcripts, indicating that Plasmax has more influence upon regulatory processes of proteins after translation. This work provides complementary information to the original study of Plasmax, aiming to facilitate the selection of appropriate media for in vitro cancer cell studies.Item Draft genome sequence of fusarium oxysporum f. sp. albedinis strain foa 133, the causal agent of bayoud disease on date palm(2020) Khayi, Slimane; Khoulassa, Siham; Gaboun, Fatima; Abdelwahd, Rabha; Diria, Ghizlane; Labhilili, Mustapha; Iraqi, Driss; El Guilli, Mohammed; Fokar, Mohamed (TTU); Mentag, RachidFusarium oxysporum f. sp. albedinis is the causal agent of vascular wilt of date palm. Here, we report the genome assembly of the Foa 133 strain, which consists of 3,325 contigs with a total length of 56,228,901 bp, a GC content of 47.42%, an N50 value of 131,587 bp, and 3,684 predicted genes.Item Genetic diversity of Moroccan date palm revealed by microsatellite markers(2023) Khoulassa, Siham; Khurshid, Haris; Fokar, Mohamed (TTU); Eliddrissy, Hicham; Outeha, Youssef; Elfadil, Jamal; Essarioui, Adil; Benlyas, Mohamed; Mentag, Rachid; Elmoualij, BenaissaThe genetic diversity between 23 Moroccan date palm cultivars collected from the National Palm Collection at the INRA (National Agricultural Research Institute) experimental field in Zagora was assessed using SSR markers that are specifically designed for date palm. Among the 16 tested SSR, 13 were successfully amplified, and were selected to carry out this study. 208 bands were amplified, ranging from 10 to 25 bands per cultivar with an average of 16 alleles per cultivar. The value of heterozygosity of the studied markers ranged from 0.11 to 0.30. The pairwise genetic distances between those cultivars ranged from 0.06 to 0.46. The hierarchical cluster analysis distributed the 23 genotypes into four different groups of one to ten cultivars.Item Glycome Profiling of Cancer Cell Lines Cultivated in Physiological and Commercial Media(2022) Wang, Junyao (TTU); Peng, Wenjing (TTU); Yu, Aiying (TTU); Fokar, Mohamed (TTU); Mechref, Yehia (TTU)A complex physiological culture medium (Plasmax) was introduced recently, composed of nutrients and metabolites at concentrations normally found in human plasma to mimic the in vivo environment for cell line cultivation. As glycosylation has been proved to be involved in cancer development, it is necessary to investigate the glycan expression changes in media with different nutrients. In this study, a breast cancer cell line, MDA-MB-231BR, and a brain cancer cell line, CRL-1620, were cultivated in Plasmax and commercial media to reveal cell line glycosylation discrepancies prompted by nutritional environments. Glycomics analyses of cell lines were performed using LC-MS/MS. The expressions of multiple fucosylated N-glycans, such as HexNAc4Hex3DeoxyHex1 and HexNAc5Hex3DeoxyHex1, derived from both cell lines exhibited a significant increase in Plasmax. Among the O-glycans, significant differences were also observed. Both cell lines cultivated in EMEM had the lowest amounts of O-glycans expressed. The original work described the development of Plasmax, which improves colony formation, and resulted in transcriptomic and metabolomic alterations of cancer cell lines, while our results indicate that Plasmax can significantly impact protein glycosylation. This study also provides information to guide the selection of media for in vitro cancer cell glycomics studies.Item Identification of Novel MeCP2 Cancer-Associated Target Genes and Post-Translational Modifications(2020) Castro-Piedras, Isabel; Vartak, David; Sharma, Monica; Pandey, Somnath; Casas, Laura; Molehin, Deborah; Rasha, Fahmida; Fokar, Mohamed (TTU); Nichols, Jacob (TTU); Almodovar, Sharilyn; Rahman, Rakhshanda Layeequr (TTU); Pruitt, KevinAbnormal regulation of DNA methylation and its readers has been associated with a wide range of cellular dysfunction. Disruption of the normal function of DNA methylation readers contributes to cancer progression, neurodevelopmental disorders, autoimmune disease and other pathologies. One reader of DNA methylation known to be especially important is MeCP2. It acts a bridge and connects DNA methylation with histone modifications and regulates many gene targets contributing to various diseases; however, much remains unknown about how it contributes to cancer malignancy. We and others previously described novel MeCP2 post-translational regulation. We set out to test the hypothesis that MeCP2 would regulate novel genes linked with tumorigenesis and that MeCP2 is subject to additional post-translational regulation not previously identified. Herein we report novel genes bound and regulated by MeCP2 through MeCP2 ChIP-seq and RNA-seq analyses in two breast cancer cell lines representing different breast cancer subtypes. Through genomics analyses, we localize MeCP2 to novel gene targets and further define the full range of gene targets within breast cancer cell lines. We also further examine the scope of clinical and pre-clinical lysine deacetylase inhibitors (KDACi) that regulate MeCP2 post-translationally. Through proteomics analyses, we identify many additional novel acetylation sites, nine of which are mutated in Rett Syndrome. Our study provides important new insight into downstream targets of MeCP2 and provide the first comprehensive map of novel sites of acetylation associated with both pre-clinical and FDA-approved KDACi used in the clinic. This report examines a critical reader of DNA methylation and has important implications for understanding MeCP2 regulation in cancer models and identifying novel molecular targets associated with epigenetic therapies.Item Metagenomics Analysis of Breast Microbiome Highlights the Abundance of Rothia Genus in Tumor Tissues(2023) Kartti, Souad; Bendani, Houda; Boumajdi, Nasma; Bouricha, El Mehdi; Zarrik, Oumaima; EL Agouri, Hajar; Fokar, Mohamed (TTU); Aghlallou, Youssef; EL Jaoudi, Rachid; Belyamani, Lahcen; Elkhannoussi, Basma; Ibrahimi, AzeddineBreast cancer is one of the main global priorities in terms of public health. It remains the most frequent cancer in women and is the leading cause of their death. The human microbiome plays various roles in maintaining health by ensuring a dynamic balance with the host or in the appearance of various pathologies including breast cancer. In this study, we performed an analysis of bacterial signature differences between tumor and adjacent tissues of breast cancer patients in Morocco. Using 16S rRNA gene sequencing, we observed that adjacent tissue contained a much higher percentage of the Gammaproteobacteria class (35.7%) while tumor tissue was characterized by a higher percentage of Bacilli and Actinobacteria classes, with about 18.8% and 17.2% average abundance, respectively. Analysis of tumor subtype revealed enrichment of genus Sphingomonodas in TNBC while Sphingomonodas was predominant in HER2. The LEfSe and the genus level heatmap analysis revealed a higher abundance of the Rothia genus in tumor tissues. The identified microbial communities can therefore serve as potential biomarkers for prognosis and diagnosis, while also helping to develop new strategies for the treatment of breast cancer patients.Item Molecular Identification of Genetic Diversity and Population Structure in Moroccan Male Date Palm (Phoenix dactylifera L.) Using Inter-Simple Sequence Repeat, Direct Amplification of Minisatellite DNA, and Simple Sequence Repeat Markers(2024) Ibrahimi, Maha; Brhadda, Najiba; Ziri, Rabea; Fokar, Mohamed (TTU); Amghar, Ilham; Gaboun, Fatima; Habach, Aicha; Meziani, Reda; Elfadile, Jamal; Abdelwahd, Rabha; Diria, GhizlaneUnderstanding genetic diversity and population structure plays a vital role in the efficient use of available material in plant-breeding programs and in germplasm conservation strategies. In the present study, we aim to evaluate the genetic variations and population structure of male date palms from Morocco. The genetic diversity of 100 date palm (Phoenix dactylifera L.) genotypes was investigated using the performance of three types of molecular markers: inter-simple sequence repeats (ISSRs), direct amplification of minisatellite DNA (DAMD), and simple sequence repeats (SSRs). On the basis of their polymorphic information content (PIC) (ISSRs = 0.38; DAMD = 0.4; SSRs = 0.33), effective multiplex ratio (EMR) (ISSRs = 27.34; DAMD = 52.31; SSRs = 22.20), Resolving power Rp (ISSR = 13.81; DAMD = 28.73; SSR = 14.6), and marker index (MI) (ISSRs = 9.22; DAMD = 20.23; SSRs = 7.54) values, all markers used in our study are considered informative markers. Among them, DAMD markers demonstrated slightly higher informativeness compared to ISSR and SSR markers. A total of 216, 438, and 248 bands were, respectively, detected using ISSRs, DAMD, and SSRs, with 95%, 98% and 94% of polymorphism, respectively. The AMOVA results revealed considerable diversity within date palms. The PCOa results showed that males of Tinghir and Errachidia were regrouped into the same cluster, while males of Goulmima were separated into another group. A cluster and structure analysis separated the studied genotypes into three groups. One group comprises genotypes of males from Zagora with some female varieties scattered in this group. The second group includes male genotypes from Goulmima along with accessions of female and male varieties. The third group contains males of Errachidia, Tata and Tinghir populations. The cluster and structure analysis separated the studied genotypes according to their origin.Item Single-nuclei analysis reveals depot-specific transcriptional heterogeneity and depot-specific cell types in adipose tissue of dairy cows(2022) Michelotti, Tainara C. (TTU); Kisby, Brent R.; Flores, Lauryn S. (TTU); Tegeler, Alexandra P. (TTU); Fokar, Mohamed (TTU); Crasto, Chiquito (TTU); Menarim, Bruno C.; Loux, Shavahn C.; Strieder-Barboza, Clarissa (TTU)Adipose tissue (AT) is an endocrine organ with a central role on whole-body energy metabolism and development of metabolic diseases. Single-cell and single-nuclei RNA sequencing (scRNA-seq and snRNA-seq, respectively) analyses in mice and human AT have revealed vast cell heterogeneity and functionally distinct subtypes that are potential therapeutic targets to metabolic disease. In periparturient dairy cows, AT goes through intensive remodeling and its dysfunction is associated with metabolic disease pathogenesis and decreased productive performance. The contributions of depot-specific cells and subtypes to the development of diseases in dairy cows remain to be studied. Our objective was to elucidate differences in cellular diversity of visceral (VAT) and subcutaneous (SAT) AT in dairy cows at the single-nuclei level. We collected matched SAT and VAT samples from three dairy cows and performed snRNA-seq analysis. We identified distinct cell types including four major mature adipocytes (AD) and three stem and progenitor cells (ASPC) subtypes, along with endothelial cells (EC), mesothelial cells (ME), immune cells, and pericytes and smooth muscle cells. All major cell types were present in both SAT and VAT, although a strong VAT-specificity was observed for ME, which were basically absent in SAT. One ASPC subtype was defined as adipogenic (PPARG+) while the other two had a fibro-adipogenic profile (PDGFRA+). We identified vascular and lymphatic EC subtypes, and different immune cell types and subtypes in both SAT and VAT, i.e., macrophages, monocytes, T cells, and natural killer cells. Not only did VAT show a greater proportion of immune cells, but these visceral immune cells had greater activation of pathways related to immune and inflammatory response, and complement cascade in comparison with SAT. There was a substantial contrast between depots for gene expression of complement cascade, which were greatly expressed by VAT cell subtypes compared to SAT, indicating a pro-inflammatory profile in VAT. Unprecedently, our study demonstrated cell-type and depot-specific heterogeneity in VAT and SAT of dairy cows. A better understanding of depot-specific molecular and cellular features of SAT and VAT will aid in the development of AT-targeted strategies to prevent and treat metabolic disease in dairy cows, especially during the periparturient period.