Induction of cell cycle arrest and apoptosis by Ormenis eriolepis a Morrocan endemic plant in various human cancer cell lines

Background: Ormenis eriolepis Coss (Asteraceae) is an endemic Moroccan subspecies, traditionally named “Hellala” or “Fergoga”. It’s usually used for its hypoglycemic effect as well as for the treatment of stomacal pain. As far as we know, there is no scientific exploration of anti tumoral activity of Ormenis eriolepis extracts.Materials and Methods: In this regard, we performed a screening of organic extracts and fractions in a panel of both hematological and solid cancer cell lines, to evaluate the potential in vitro anti tumoral activity and to elucidate the respective mechanisms that may be responsible for growth arrest and cell death induction. The plant was extracted using organic solvents, and four different extracts were screened on Jurkat, Jeko-1, TK-6, LN229, SW620, U2OS, PC-3 and NIH3T3 cells.Results: Cell viability assays revealed that, the IC50 values were (11,63±5,37μg/ml) for Jurkat, (13,33±1,67μg/ml) for Jeko-1, (41,67±1,98μg/ml) for LN229 and (19,31±4,88μg/ml) for PC-3 cells upon treatment with Oe-DF and Oe-HE respectively. Both the fraction and extract exhibited no effects on TK6 and NIH3T3. Cytometry analysis accompanied by DNA damage signaling protein levels monitoring (p-H2A.X), showed that both the Dichloromethane Fraction and Hexanic extract induce DNA double stranded breaks (DSBs) accompanied by cell cycle arrest in G1 (Jurkat, Jeko -1 and LN22) and G2/M (PC-3) phases which is agreed with the caspase activity observed. Additional experiments with selective inhibitors of stress and survival pathways (JNK, MAPK, Rho, p53, and JAK3) indicated that none of these pathways was significantly involved in apoptosis induction. The bioactive compound analysis by CG/MS indicated that the major compounds in Oe-DF were: Linoleic Acid (15,89%), Podophyllotoxin (17,89%) and Quercetin (22,95%). For Oe-HE the major molecules were: Linoleic Acid (9,76%), α-curcumene (7,07%), α-bisabolol (5,49%), Campesterol (4,41%), Stigmasterol (14,08%) and β-sitosterol (7,49%).Conclusion: Our data suggest that bioactive compounds present in Ormenis eriolepis show significant anti proliferative activity inducing cell cycle arrest and cell death operating through apoptosis pathway.Keywords: Ormenis eriolepis Coss, cancer cell lines, cytotoxicity, apoptosis, bioactive compound

Identification of single nucleotide variants in the Moroccan population by whole-genome sequencing

Background: Large-scale human sequencing projects have described around a hundred-million single nucleotide variants (SNVs). These studies have predominately involved individuals with European ancestry despite the fact that genetic diversity is expected to be highest in Africa where Homo sapiens evolved and has maintained a large population for the longest time. The African Genome Variation Project examined several African populations but these were all located south of the Sahara. Morocco is on the northwest coast of Africa and mostly lies north of the Sahara, which makes it very attractive for studying genetic diversity. The ancestry of present-day Moroccans is unknown and may be substantially different from Africans found South of the Sahara desert, Recent genomic data of Taforalt individuals in Eastern Morocco revealed 15,000-year-old modern humans and suggested that North African individuals may be genetically distinct from previously studied African populations. Results: We present SNVs discovered by whole genome sequencing (WGS) of three Moroccans. From a total of 5.9 million SNVs detected, over 200,000 were not identified by 1000G and were not in the extensive gnomAD database. We summarise the SNVs by genomic position, type of sequence gene context and effect on proteins encoded by the sequence. Analysis of the overall genomic information of the Moroccan individuals to individuals from 1000G supports the Moroccan population being distinct from both sub-Saharan African and European populations. Conclusions: We conclude that Moroccan samples are genetically distinct and lie in the middle of the previously observed cline between populations of European and African ancestry. WGS of Moroccan individuals can identify a large number of novel SNVs and aid in functional characterisation of the genome

The evolving SARS-CoV-2 epidemic in Africa: Insights from rapidly expanding genomic surveillance

INTRODUCTION Investment in Africa over the past year with regard to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) sequencing has led to a massive increase in the number of sequences, which, to date, exceeds 100,000 sequences generated to track the pandemic on the continent. These sequences have profoundly affected how public health officials in Africa have navigated the COVID-19 pandemic. RATIONALE We demonstrate how the first 100,000 SARS-CoV-2 sequences from Africa have helped monitor the epidemic on the continent, how genomic surveillance expanded over the course of the pandemic, and how we adapted our sequencing methods to deal with an evolving virus. Finally, we also examine how viral lineages have spread across the continent in a phylogeographic framework to gain insights into the underlying temporal and spatial transmission dynamics for several variants of concern (VOCs). RESULTS Our results indicate that the number of countries in Africa that can sequence the virus within their own borders is growing and that this is coupled with a shorter turnaround time from the time of sampling to sequence submission. Ongoing evolution necessitated the continual updating of primer sets, and, as a result, eight primer sets were designed in tandem with viral evolution and used to ensure effective sequencing of the virus. The pandemic unfolded through multiple waves of infection that were each driven by distinct genetic lineages, with B.1-like ancestral strains associated with the first pandemic wave of infections in 2020. Successive waves on the continent were fueled by different VOCs, with Alpha and Beta cocirculating in distinct spatial patterns during the second wave and Delta and Omicron affecting the whole continent during the third and fourth waves, respectively. Phylogeographic reconstruction points toward distinct differences in viral importation and exportation patterns associated with the Alpha, Beta, Delta, and Omicron variants and subvariants, when considering both Africa versus the rest of the world and viral dissemination within the continent. Our epidemiological and phylogenetic inferences therefore underscore the heterogeneous nature of the pandemic on the continent and highlight key insights and challenges, for instance, recognizing the limitations of low testing proportions. We also highlight the early warning capacity that genomic surveillance in Africa has had for the rest of the world with the detection of new lineages and variants, the most recent being the characterization of various Omicron subvariants. CONCLUSION Sustained investment for diagnostics and genomic surveillance in Africa is needed as the virus continues to evolve. This is important not only to help combat SARS-CoV-2 on the continent but also because it can be used as a platform to help address the many emerging and reemerging infectious disease threats in Africa. In particular, capacity building for local sequencing within countries or within the continent should be prioritized because this is generally associated with shorter turnaround times, providing the most benefit to local public health authorities tasked with pandemic response and mitigation and allowing for the fastest reaction to localized outbreaks. These investments are crucial for pandemic preparedness and response and will serve the health of the continent well into the 21st century

Gut-Lung Axis in COVID-19

COVID-19 is a pandemic infection of the respiratory system caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The viral ribonucleic acid (RNA) was found in many parts of the COVID-19 patients including the stool, suggesting a potential interaction with the host’s gut microbiome. The gut microbiome also plays major roles in immunity and inflammation. It also impacts pulmonary functions through the gut-lung axis. There have been recent reports of the importance of the host microbiome in infection and pathogenicity. The understanding of the gut and lung microbiomes would open the gate to new therapeutic approaches

Human Immunodeficiency Virus Type 1 Inhibitory Activity of

Extracts from a new chemotype of Mentha longifolia, a mint species that grows spontaneously and widely in the Moroccan mountains, were tested against human immunodeficiency virus type 1 (HIV-1). We observed that non-toxic concentrations (10 µg/mL) of extracts from this plant, in particular methanol (Ext-1) and ethyl acetate (Ext-3) extracts, significantly inhibit (p < 0.01) HIV-1BaL infection by about 40% and 55%, respectively. In addition, only Ext-3 shows significant (p < 0.008) inhibitory activity (50% inhibition) against HIV-1 reverse transcriptase. It is noteworthy that chemical analysis of these extracts suggests that flavonoids, mainly flavones of M. longifolia, may be the major inhibitors of HIV infection. In conclusion, these in vitro data suggest that components of M. longifolia may represent potential anti-HIV agents; the identification of such components is in progress

Retama monosperma n-hexane extract induces cell cycle arrest and extrinsic pathway-dependent apoptosis in Jurkat cells

BACKGROUND: Retama monosperma L. (Boiss.) or Genista monosperma L. (Lam.), locally named as “R’tam”, is an annual and spontaneous plant belonging to the Fabaceae family. In Morocco, Retama genus is located in desert regions and across the Middle Atlas and it has been widely used in traditional medicine in many countries. In this study, we show that Retama monosperma hexane extract presents significant anti-leukemic effects against human Jurkat cells. METHODS: Human Jurkat cells, together with other cell lines were screened with different concentrations of Retama monosperma hexane extract at different time intervals. Growth inhibition was determined using luminescent-based viability assays. Cell cycle arrest and apoptosis were measured by flow cytometry analysis. Combined caspase 3 and 7 activities were measured using luminometric caspase assays and immunoblots were performed to analyze expression of relevant pro- and anti-apoptotic proteins. GC-MS were used to determine the chemical constituents of the active extract. RESULTS: Retama monosperma hexane extract (Rm-HE) showed significant cytotoxicity against Jurkat cells, whereas it proved to be essentially ineffective against both normal mouse fibroblasts (NIH3T3) and normal lymphocytes (TK-6). Cytometric analysis indicated that Rm-HE promoted cell cycle arrest and apoptosis induction accompanied by DNA damage induction indicated by an increase in p-H2A.X levels. Rm-HE induced apoptosis was partially JNK-dependent and characterized by an increase in Fas-L levels together with activation of caspases 8, 3, 7 and 9, whereas neither the pro-apoptotic nor anti-apoptotic mitochondrial membrane proteins analyzed were significantly altered. Chemical identification analysis indicated that α-linolenic acid, campesterol, stigmasterol and sitosterol were the major bioactive components within the extract. CONCLUSIONS: Our data suggest that bioactive compounds present in Rm-HE show significant anti leukemic activity inducing cell cycle arrest and cell death that operates, at least partially, through the extrinsic apoptosis pathway

Gut microbiome compositional differences between tumor and Non-tumor adjacent tissues from US and Spain identifies eikenella as a potential new player in colorectal cancer

Trabajo presentado en el 5th International Human Microbiome Congress, IHMC, celebrado en Luxemburgo del 31 de marzo al 2 de abril de 2015Colorectal cancer (CRC) is the third most common cancer in the world and the second leading cause of cancer deaths in the US and Spain. The molecular mechanisms involved in the etiology of CRC are not yet elucidated due in part to the complexity of the human gut microbiota. In this study, we compared the microbiome composition of 102 tumor and matching adjacent normal tissue (normal) from cohorts from the US and Spain by 16S rRNA amplicon sequencing in order to determine the impact of the geographic origin on the CRC microbiome. Data showed a significantly (P < 0.05) higher Phylogenetic Diversity (PD) for the US (PD Normal = 26.3 ± 5.3, PD Tumor = 23.3 ± 6.2) compared to the Spanish cohort (PD Normal = 18.9 ± 5.9, PD Tumor = 18.7 ± 6.6) while no significant differences in bacterial diversity were observed between tumor and normal tissues for individuals from the same country. Normal tissues from the Spanish cohort were enriched in Firmicutes (43.9% and 22.2%, P = 0.0001) and Actinobacteria (1.6% and 0.5%, P = 0.0018) compared to US normal tissues, while normal tissues from the US had significantly higher abundances of Fusobacteria (8.1% and 1.5%, P = 0.0023) and Sinergistetes (0.3% and 0.1%, P = 0.0097). Comparison of tumor and normal tissues in each cohort identified the genus Eikenella significantly over represented in US tumors (0.024% and 0%, P = 0.03), and the genera Fusobacterium (10.4% and 1.5%, P = < 0.0001), Bulleida (0.36% and 0.09%, P = 0.02), Gemella (1.46% and 0.19%, P = 0.03), Parvimonas (3.14% and 0.86%, P = 0.03), Campylobacter (0.15% and 0.008%, P = 0.047), and Streptococcus (2.84% and 2.19%, P = 0.05) significantly over represented in Spanish tumors. Our study suggests that microbiome compositional dissimilarities by geographic location should be taken in consideration when approaching CRC therapeutic options.Peer reviewe

Preclinical Evaluation of panobinostat and ONC201 for the treatment of diffuse intrinsic pontine glioma (DIPG)

Diffuse intrinsic pontine glioma (DIPG) also referred as paediatric high-grade glioma (pHGG) is a fast-growing and aggressive type of childhood brain cancer. Recent studies investigating the molecular pathogenesis of DIPG have identified new therapeutic targets, paving the way for a new line of drugs mainly HDAC inhibitors. However, despite long years of trials, no significant results have been generated yet. Panobinostat is a HDAC inhibitor that has shown promising preclinical cytotoxicity in DIPG but failed so far in clinical trials. This study aims to re-evaluate the efficacy of Panobinostat in DIPG in vitro using patient-derived DIPG cell cultures obtained directly from patients. ONC201 is another potentially effective drug in DIPG. This apoptotic agent has been considered in a few clinical trials in diffuse glioma including DIPG. Our results reveal a dose-dependent response to Panobinostat and ONC201 in DIPG cells. However, Panobinostat caused a significant reduction in the mean percentage cell viability at a lower concentration compared to ONC201. Panobinostat caused significant decreases in DIPG cell viability at concentrations greater than or equal to 0.002 μM (p<0.05), the response reached a plateau after 0.1 μM, which reduced cell viability to 32.81 % ± 0.25 % (p = 6.74E−06) when compared to control cells. ONC201 only significantly induced apoptosis at concentrations equal or higher than 0.01 μM (p<0.05), with its effect plateauing after 0.2 μM. This pre-clinical study supports the effectiveness of Panobinostat as a potential therapeutic agent for DIPG compared to ONC201, with no apparent synergistic effect observed in combination

Diferencias en el microbioma asociado a tejido tumoral y tejido sano adyacente en pacientes con cáncer de colon en España y Estados Unidos

Trabajo presentado en el VII Workshop Probióticos, Prebióticos y Salud: Evidencia Científica, celebrado en Sevilla (España) el 28 y 29 de enero de 2016El cáncer colorectal (CCR) es el tercero más común en el mundo y la segunda causa de muerte por cáncer en Estados Unidos (EU) y España. En los últimos años se han llevado a cabo diversos estudios del microbioma intestinal en esta patología, en la mayor parte de los casos con muestras de heces. Se ha observado alteraciones en la microbiota o disbiosis asociada al CRC y se ha propuesto un modelo microbiano para el desarrollo de inflamación crónica en la que estaría involucrada la microbiota intestinal. En este trabajo se presenta la comparación del microbioma, (composición microbiana y función inferida) determinado mediante secuenciación masiva y análisis bioinformático de amplicones del ARNr 16S, de tejido intestinal (sano y tumoral) obtenido durante recesiones de colon en pacientes con CRC en EU y España. En cuanto a la comparación entre tejido tumoral y adyacente se observaron diferencias significativas en la diversidad bacteriana para los individuos de un mismo país, con una menor riqueza de especies en los tumores. La comparación entre ambos tipos de tejido en cada cohorte identificó posibles patógenos, como Eikenella, sobrerrepresentados en los tumores de EU, mientras que Fusobacterium, y Campylobacter fueron significativamente más relevantes en los tumores españoles. La predicción funcional reveló perfiles funcionales parecidos pero con rutas metabólicas y enzimas diferencialmente representados en los tejidos tumorales. En general los datos mostraron diferencias composicionales entre ambas cohortes de estudio que deben de tenerse en cuenta previamente a abordar cualquier estudio del microbioma de CRC.Peer reviewe

Antiproliferative Effects of Withanolides from

Extracts of Withania adpressa Coss. (Solanaceae), a medicinal plant endemic to Moroccan Sahara, were tested for their cytotoxicity towards a panel of cancer cell lines (Hep2, HT29, RD, Vero and MDCK), using the (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) [MTT assay, Sigma-Aldrich]. The bioassay-guided fractionation of this plant extracts results a novel withanolide 14α,15α,17$\beta$,20β-tetrahydroxy-1-oxo-(22R)-witha-2,5,24-trienolide and the already identified withanolides F and J extract, semi-purified fractions and pure compounds exhibits potent cytotoxicity against human cancer cell lines tested, in dose-dependant manner. Morphological features of treated Hep2 cells with the novel withanolide and characteristic DNA fragmentation revealed that the cytotoxicity was due to induction of apoptosis. Taken together, the results suggest that withanolides from W. adpressa Coss. hold potential as antiproliferative agents