Case Study Of DLBCA

MiR-21 has been found to be over expressed in 20 pancreatic carcinoma tissues and cell lines compared to normal tissue or cell lines. (15)

Recent reports indicate the potential role of miRNAs as both therapeutic targets and biomarkers in IPF (5). MiRNAs are small (approximately 22 nucleotides in length) and stable non-coding molecules, which can regulate gene expression (6). They are important regulators of physiological processes, such as cellular proliferation leading to lung tissue repair. The miR200 family is functionally involved in canonical pathways of immune response and cell-cell communication. Here we propose novel studies using miR200b and miR200c in IPF compared to controls, first, to determine their function in the niche composed of ATII cells and fibroblasts, second, analyze their potential to stimulate ATII cell proliferation and transdifferentation to alveolar type I cells, third to analyze their capacity to inhibit the growth of fibroblasts; and fourth, to use them as early biomarkers of IPF development. The rationale for using 200miRNA family members as curing IPF drug is based on their dysregulated expression observed in IPF (7). Moreover, our

Deciphering MECP2-associated disorders: disrupted circuits and the hope for repair). Recently, Mellios and collaborators identified novel MECP2-regulated miRNAs during the human prenatal neurodevelopment. Utilizing human patient- and MeCP2 knockdown-derived monolayer neuronal culture and organoids models, they found an upregulation of the expression levels of miR-199 and miR-214 and their downstream targets (PAK4 and PTEN, respectively), that differentially regulate extracellular signal regulated kinase (ERK) and mitogen-activated kinase and protein kinase B (PKB/AKT) signalling pathways upstream of neurogenesis. The miR199/214 overexpression was correlated with aberrant neurogenesis and neuronal migration, whereas the inhibition of their expression rescues these effects, suggesting that the neurogenesis may be impacted in RTT via MeCP2-regulated miRNAs that acts as important effectors of neuronal development modulating molecular pathways involved in proliferation and differentiation (Mellios N et al, MeCP2-regulated miRNAs control early human neurogenesis through differential effects on ERK and AKT signaling. Molecular Psychiatry (2017) 00,

microRNAs (miRNAs) are a short ( about 22-26 nt length ) non coding RNA that involved in regulating gene expression at the level of post-transcription or translation and apparently control a broad range of processes of multicellular organisms including cellular differentiation, proliferation and apoptosis via targeting mRNA and cause their degradation or protein translation inhibition. miRNA transcribed from long pri-miRNA in the nucleus then processed by Dicer enzyme in the cytoplasm to form mature miRNA that incorporate in RNA induced silencing complex (RISC) to enhance their functions.

Specific Aim #1. To determine the effect of miR200 family on human primary ATII cell biology and lung regeneration in IPF.

In addition to transcriptional regulation and epigenetic mechanisms, posttranscriptional regulation by microRNAs is also critically involved in the intracellular signaling of oligodendrocyte differentiation. Studies using transgenic mice in which microRNA processing is specifically disrupted in the oligodendrocyte lineage by way of Dicer enzyme knockout have shown that microRNA processing is indispensible for normal CNS myelination (Dugas et al., 2010), and that in vitro OPCs also fail to differentiate in the absence of mature microRNAs. Specifically, three microRNAs including miR-219, miR-138, and miR-338 are found to be elevated by 1-2 orders of magnitude during OPC differentiation into oligodendrocytes, and that miR-219 when induced alone, is sufficient to promote the differentiation (Dugas et al., 2010). The downstream target of miR-219, ELOVL7, is also identified as a main molecular component involved in the development of the Dicer mutant phenotype (Shin, Shin, McManus, Ptacek, & Fu, 2009), and that overexpression of ELOVL7 results in lipid accumulation, which is in turn suppressed by co-overexpression of miR-219. Other important target genes of the above mentioned microRNAs include PDGFRa, Sox6, and Hes5 (Dugas et al., 2010; X. Zhao et al., 2010), all of which serve to

Aim 1.3 – Determine and evaluate the oncogenic lncRNA landscape with validated activity in the neural tube.

Aim1.1 Testing miR-125a overexpression can be performed via transforming a retrovirus vector construct into E.coli plasmid to amplify the vector, following by isolation of plasmids using specific Mini-prep Kit. Then, plasmids are transfected into Human Embryonic Kidney 293 cell line (HEK293 cell) or CH12F3 cell line. To determine the efficiency of transfection, we monitored green fluorescent protein (GFP) reporter construct expression by conducting either immunofluorescent assay or fluorescence-activated cell sorting (FACS) 14. We transduce the virus containing miR-125a vector into primary B cells that is purified by mouse B cell isolation Kit and stimulated primarily with LPS followed by second stimulation using LPS and IL-4. Cells were collected 60 h later for qRT-PCR and RNA extraction, or 96 h later for surface Ig analysis by FACS. The stimulated B cells are stained with CD19 and CD138 to measure the percentages of switched B cells by FACS. We predict a reduction in IgG1, IgG3, and CD138 populations due to the impede function in CSR and plasma cell differentiation due to miR-125a expression, which target Aicda and Prdm1. In addition, RNA extraction will be

To accomplish this, researchers knocked down Malat1 expression using short hairpin RNA (shRNA) molecules, which resulted in elevated levels of myogenic RNA and protein markers2. Therefore, the reduction in Malat1 expression following shRNA knockdown allowed for increased myogenic protein synthesis. Consequently, Malat1 expression suppressed C2C12 differentiation and promoted cellular proliferation2. Additionally, an RNA immunoprecipitation assay (RIP) was used to study the mechanism of interaction between Malat1 ncRNA and the repressive cofactor Suv39h12. Researchers utilized antibodies with an affinity for Suv39h1 to precipitate the cofactor out of the C2C12 myoblasts2. Once the Suv39h1 complexes were precipitated, the cellular material was purified and analyzed using reverse transcription polymerase chain reaction (RT-PCR)2. Following RT-PCR analysis, researchers found significant quantities of Malat1 associated with the precipitated antibody/cofactor complex, indicating that Malat1 interacted with Suv39h1to repress the MyoD gene loci in the differentiating

Analysis of adipose derived induced pluripotent stem cells (iPSC’s) microRNA (miRNA) expression profile over multiple passages and pancreatic β-cell miRNA expression profile post differentiation from iPSC’s.

Exome sequencing is the most widely used targeted sequencing method. The exome only represents less than 2% of the genetic code, but contains about 85% of known disease-related variants, making a cost-effective alternative to whole-genome sequencing2. Exome sequencing detects variants in coding exons, with the capability to expand targeted content to include untranslated regions and microRNA for a more comprehensive view of gene regulation3. DNA libraries can be prepared in one day and require only 4–5 Gb of sequencing per exome4. Exome sequencing enables researchers to focus their resources on the genes most likely to affect phenotype, it has a wide range of applications, including population genetics, genetic disease, and cancer studies3.

MicroRNAs (miRNAs) are short non-coding RNAs that regulate gene expression in plants and animals (Brennecke et al, 2005). They consist of approximately 20 to 24 nucleotides of RNA. One process by which miRNAs inhibit gene expression occurs through translational repression (Gu and Kay, 2010). Further to translational repression, endonuclease cleavage can also help repression to occur. According to Cheng et al (2015) ‘dysregulated microRNA expression is correlated with many human diseases’. This suggests that miRNAs can be used as a biomarker for cancer and some diseases such as Cardiovascular Disease (CVD). This is one of the reasons why the scientific community is interested in studying these molecules.

2009, Smalheiser NR, Neuromolecular Med. 2009, Ye Y Neural Plast. 2016, Kerstin Weiß European Journal of Cell Biology 2015, Neil R. Smalheiser Phil. Trans. R. Soc. B 2014, Iyengar BR Front Cell Neurosci. 2014). Certain specific miRNAs, for instance, miR-134, has been found in the dendritic spines of cultured hippocampal neurons (Schratt et al. 2006). It appears that these miRNAs are engaged in activity dependent, local modulation of translation that can in turn lead to stimulus dependent remodeling of dendritic spines. It is becoming increasingly obvious that many steps of gene expression, especially miRNA biogenesis and splicing, conventionally thought to be performed in the nucleus also occur in the dendritic spine. The presence of mRNAs that contain retained introns and precursors of miRNAs in the dendritic spines (Lugli et al. 2008) suggest that RNA is held in an inactive state which can be rapidly processed following external stimuli. Apart from miRNAs, the lncRNA being multifunctional molecule have emerged as an important component in the specialized neuronal compartments.

MicroRNAs (miRNAs) comprise a large family, its having approximately 20–22 nucleotides, and it regulates the expression of target genes at the post-transcriptional level. However, thousands of miRNAs have been identified in many organisms (roundworms, flies, fish, frogs, mammals, flowering plants, mosses, and even viruses) by using genetics, molecular cloning and predictions from bioinformatics technique (Lagos et al., 2001; Lim et al., 2003; Pfeffer et al., 2004; Axtell and Bartel, 2005). The whole human genome encodes at least 474 miRNA genes (Griffiths, 2004; Griffiths et al., 2006), and in human breast cancer there are 133 miRNAs were identified. MiRNAs control messenger RNA expression and which are frequently dysregulated in

Currently, I am involved in developing methodology to identifying differentially expressed genes using single cell RNA sequencing data. scRNA-Seq data has tremendous opportunity to study the expressions of genes at an individual cellular level. However, this type of