Human mesenchymal stem cells-mediated transcriptomic regulation of leukemic cells in delivering anti-tumorigenic effects

Treatment of leukemia has become much difficult because of resistance to the existing anticancer therapies. This has thus expedited the search for alternativ therapies, and one of these is the exploitation of mesenchymal stem cells (MSCs) towards control of tumor cells. The present study investigated the effect of human umbilical cord-derived MSCs (UC-MSCs) on the proliferation of leukemic cells and gauged the transcriptomic modulation and the signaling pathways potentially affected by UC-MSCs. The inhibition of growth of leukemic tumor cell lines was assessed by proliferation assays, apoptosis and cell cycle analysis. BV173 and HL-60 cells were further analyzed using microarray gene expression profiling. The microarray results were validated by RT-qPCR and western blot assay for the corresponding expression of genes and proteins. The UC-MSCs attenuated leukemic cell viability and proliferation in a dose-dependent manner without inducing apoptosis. Cell cycle analysis revealed that the growth of tumor cells was arrested at the G0/G1 phase. The microarray results identified that HL-60 and BV173 share 35 differentially expressed genes (DEGs) (same expression direction) in the presence of UC-MSCs. In silico analysis of these selected DEGs indicated a significant influence in the cell cycle and cell cycle-related biological processes and signaling pathways. Among these, the expression of DBF4, MDM2, CCNE2, CDK6, CDKN1A, and CDKN2A was implicated in six different signaling pathways that play a pivotal role in the anti-tumorigenic activity exerted by UC-MSCs. The UC-MSCs perturbate the cell cycle process of leukemic cells via dysregulation of tumor suppressor and oncogene expression

Interleukin-21 gene polymorphisms in Iranian multiple sclerosis subjects

Multiple Sclerosis (MS) as an autoimmune inflammatory disease which affects the central nervous system (CNS), is reportedly the second most widespread culprit for neurological disability among young adults. Ranging between 2 and 150 per 100,000, MS is prevalent everywhere in the world. In recent years, the increasing incidence of MS in Iran has been a matter of interest for investigators. Even though MS cannot be claimed to be directly hereditary, genetics variants are reportedly related with MS. Cytokines play an important role in the initiation and maintenance of autoimmune diseases including MS. As a new pro-inflammatory cytokine, interleukin-21 (IL-21) has recently been observed as a target of intervention in many autoimmune diseases such as MS. On the other hand, IL-21 is a cytokine that is produced by T helper 17 (Th17) cells and functions as a growth factor for Th17 cells, considering the importance of IL-21 in Th17 cells development as a new central pathogen in MS disease also new finding on the role of IL-21 in MS disease, association of IL-21 gene polymorphisms (C5250T and G1472T) with MS disease in Iranian subjects were investigated. In this study 301 Iranian MS patients (224 females and 77 males) with mean age of 33.26 +/- 8.74 Years and 201 healthy controls (149 female and 52 male) with mean age of 32.4 +/-8.64 were used to determine the G1472T polymorphism in the second intron (rs2055979) and C5250T polymorphisms in the Exone 3 (rs4833837) of IL- 21 gene. Among patients under study, information related to disease type, EDSS score, Progression Index and age of disease onset were obtained. IL-21 G1472T polymorphism was identified by RFLP-PCR method while the IL-21 C5250T polymorphism was genotyped via allele specific oligonucleotide PCR (ASO-PCR). Genotype and allele frequencies were compared between patients and controls by chi-square. Association of polymorphisms with disease types, age at disease onset, EDSS score and progression index were also investigated. Genotype and Allele frequencies at +1472 G/T position were compared between normal population and patients group. On evaluation of genotypes and allele frequency at this position, there was a significant difference between cases and controls (p=0.003). Also, association of type of disease with G1472T polymorphism was analyzed and a significant difference in genotype frequency was observed between patients and controls group (p=0.0001). The association of G1472T polymorphism with EDSS, progression index and onset age were not significant (p=0.8, p=0.35, p=0.16, respectively). At +5250 C/T position among 301 patients under study, genotype distribution was determined Differences between cases and controls was not significant (p=0.95). Of the 310 patients studied at +5250 C/T position, type disease was determined in 223 patients. Statistical analysis showed no significant difference between genotype frequency and disease type (p=0.71). The association of C5250T with progression index, EDSS and age at disease onset, were not significant (p=0.8, p=0.35 and p=0.16, respectively). In conclusion this study showed that IL-21 gene polymorphism (G1472T) was significantly associated with MS development. Other studies to clarify the association of this polymorphism with MS susceptibility in other populations would be desired

Expression profiling of diabetic and hypertension-associated genes of Malaysian male subjects with coronary heart disease in a hospital in Malaysia

Coronary Heart Disease (CHD) is still the number-one killer in the world. As for Malaysia, the number of people with CHD has more than tripled in the past 40 years and the figures are still growing. Notably, many of the patients with CHD have at least one modifiable risk factor such as hypertension (HT) and diabetes mellitus (DM). HT and DM stand out as two major modifiable risk factors for CHD and it is well established that the incidence of over 80% of CHD is attributable to these two modifiable risk factors. This study was carried out for the purpose of profiling expression of DM and HT associated genes and identify related biological process and modulated signaling pathways of Malaysian male subjects with CHD from three ethnic group, namely Malay, Chinese and Indian. In order to achieve the goal in group A, four groups of subjects were divided into: 1) 42 healthy subjects; 2) 42 subjects with only DM; 3) 42 subjects with only CHD, and 4) 42 subjects with CHD + DM. The RNA was extracted from blood specimens by mean of commercial extraction kits. The RT2 Profiler™ PCR Array was utilized to determine gene profiling on group 1 and group 2, group 1 and group 3, group 1 and group 4. To validate the results of RT2 profiler™ PCR Array, three of significantly dysregulated genes were selected and validation was conducted through Q-RT-PCR in a larger and independent population. For this purpose, new subjects were divided into: 1) 75 healthy subjects. 2) 75 subjects with DM+CHD. The Same pattern was followed in group B, DM replaced by HT in the groups with the same numbers, to investigate and identify risk genes and modulated pathway/s related to HT which confer risk to CHD development. In order to validate the result in group B, single gene expression profiling was performed between 75 healthy individuals and 75 patients suffering from HT+CHD through Q-RT-PCR (An independent and larger population). In group A, 12 significantly dysregulated genes related to diabetes and Toll-Like receptor signaling pathway were identified which may be a culprit to susceptible diabetic patients to CHD development. In Silico experiments imply a role for inflammatory responses in the circulating leukocytes as a biomarker reflecting initiation of CHD in patients with DM. In group B, 16 significantly dysregulated genes related to hypertension were identified and the RAAS cell signaling pathway was highlighted as a culprit in people suffering from hypertension which may be prone to CHD. In Silico analysis showed that the majority of the identified genes involved in renin-angiotensin regulation and other categories related to renin-angiotensin such as, regulation of blood volume and regulation of blood vessel by renin-angiotensin. In conclusion, some differentially dysregulated genes and modulated pathways were identified which warrant further investigation in the setting of CHD and its risk factors. It is hoped that a greater understanding of genetic predisposition to CHD will unravel clues to its etiology and allow development of novel diagnostic and therapeutic tools to permit targeted interventions to reduce this global health burden

Genetic analysis of the atrial natriuretic peptide gene polymorphisms among essential hypertensive patients in Malaysia

Background. Atrial natriuretic peptide (ANP) considerably influences blood pressure regulation through water and sodium homoeostasis. Several of the studies have utilized anonymous genetic polymorphic markers and made inconsequent claims about the ANP relevant disorders. Thus, we screened Insertion/Deletion (ID) and G191A polymorphisms of ANP to discover sequence variations with potential functional significance and to specify the linkage disequilibrium pattern between polymorphisms. The relationships of detected polymorphisms with EH with or without Type 2 Diabetes Mellitus (T2DM) status were tested subsequently. Method. ANP gene polymorphisms (I/D and A191G) were specified utilizing mutagenically separated Polymerase Chain Reaction (PCR) in 320 subjects including 163 EH case subjects and 157 controls. Result. This case-control study discovered a significant association between I/D polymorphisms of ANP gene in EH patient without T2DM. However, the study determined no association between G191A polymorphisms of ANP in EH with or without T2DM. In addition, sociodemographic factors in the case and healthy subjects exhibited strong differences (). Conclusion. As a risk factor, ANP gene polymorphisms may affect hypertension. Despite the small sample size in this study, it is the first research assessing the ANP gene polymorphisms in both EH and T2DM patients among Malaysian population

Expression Profiling of Genes Related to Endothelial Cells Biology in Patients with Type 2 Diabetes and Patients with Prediabetes

Endothelial dysfunction appears to be an early sign indicating vascular damage and predicts the progression of atherosclerosis and cardiovascular disorders. Extensive clinical and experimental evidence suggests that endothelial dysfunction occurs in Type 2 Diabetes Mellitus (T2DM) and prediabetes patients. This study was carried out with an aim to appraise the expression levels in the peripheral blood of 84 genes related to endothelial cells biology in patients with diagnosed T2DM or prediabetes, trying to identify new genes whose expression might be changed under these pathological conditions. The study covered a total of 45 participants. The participants were divided into three groups: group 1, patients with T2DM; group 2, patients with prediabetes; group 3, control group. The gene expression analysis was performed using the Endothelial Cell Biology RT2 Profiler PCR Array. In the case of T2DM, 59 genes were found to be upregulated, and four genes were observed to be downregulated. In prediabetes patients, increased expression was observed for 49 genes, with two downregulated genes observed. Our results indicate that diabetic and prediabetic conditions change the expression levels of genes related to endothelial cells biology and, consequently, may increase the risk for occurrence of endothelial dysfunction

Signature pattern of gene expression and signaling pathway in premature diabetic patients uncover their correlation to early age Coronary Heart Disease

Background: Coronary Heart Disease (CHD) is the leading cause of death in industrialized countries. There is currently no direct relation between CHD and type 2 diabetes mellitus (T2D), one of the major modifiable risk factors for CHD. This study was carried out for genes expression profiling of T2D associated genes to identify related biological processes/es and modulated signaling pathway/s of male subjects with CHD. Method: the subjects were divided into four groups based on their disease, including control, type 2 diabetes mellitus (T2D), CHD, and CHD + T2D groups. The RNA was extracted from their blood, and RT2 Profiler™ PCR Array was utilized to determine gene profiling between groups. Finally, the PCR Array results were validated by using Q-RT-PCR in a more extensive and independent population. Result: PCR Array results revealed that the T2D and T2D + CHD groups shared 11 genes significantly up-regulated in both groups. Further analysis showed that the mRNA levels of AKT2, IL12B, IL6, IRS1, IRS2, MAPK14, and NFKB1 increased. Consequently, the mRNA levels of AQP2, FOXP3, G6PD, and PIK3R1 declined in the T2D + CHD group compared to the T2D group. Furthermore, in silico analysis indicated 36 Gene Ontology terms and 59 signaling pathways were significantly enriched in both groups, which may be a culprit in susceptibility of diabetic patients to CHD development. Conclusion: Finally, the results revealed six genes as a hub gene in altering various biological processes and signaling pathways. The expression trend of these identified genes might be used as potential markers and diagnostic tools for the early identification of the vulnerability of T2D patients to develop premature CHD

Signature pattern of gene expression and signaling pathway in premature diabetic patients uncover their correlation to early age coronary heart disease

Background Coronary Heart Disease (CHD) is the leading cause of death in industrialized countries. There is currently no direct relation between CHD and type 2 diabetes mellitus (T2D), one of the major modifiable risk factors for CHD. This study was carried out for genes expression profiling of T2D associated genes to identify related biological processes/es and modulated signaling pathway/s of male subjects with CHD. Method the subjects were divided into four groups based on their disease, including control, type 2 diabetes mellitus (T2D), CHD, and CHD + T2D groups. The RNA was extracted from their blood, and RT2 Profiler™ PCR Array was utilized to determine gene profiling between groups. Finally, the PCR Array results were validated by using Q-RT-PCR in a more extensive and independent population. Result PCR Array results revealed that the T2D and T2D + CHD groups shared 11 genes significantly up-regulated in both groups. Further analysis showed that the mRNA levels of AKT2, IL12B, IL6, IRS1, IRS2, MAPK14, and NFKB1 increased. Consequently, the mRNA levels of AQP2, FOXP3, G6PD, and PIK3R1 declined in the T2D + CHD group compared to the T2D group. Furthermore, in silico analysis indicated 36 Gene Ontology terms and 59 signaling pathways were significantly enriched in both groups, which may be a culprit in susceptibility of diabetic patients to CHD development. Conclusion Finally, the results revealed six genes as a hub gene in altering various biological processes and signaling pathways. The expression trend of these identified genes might be used as potential markers and diagnostic tools for the early identification of the vulnerability of T2D patients to develop premature CHD. Graphical AbstractApplied Science, Faculty ofNon UBCBiomedical Engineering, School ofReviewedFacultyResearche