Farinaz Afsari, PhD

 I was a Faculty, Author and Instructor in Biology at PEOI: Professional Educational Organisation International until 2021.

During my work at PEOI, I was constructing courses in " Cell Signalling Pathways" for students nationally and internationally who had financial and other difficulties, hindering their ability to attend universities.

 

 I was awarded a Certificate for the work I did at PEOI as a part of UNV on 29th of June 2020.

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« Professional Education, Testing and Certification International Fund awards this certificate in recognition of your contribution to the Sustainable Development Goals in PEOI »

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In past, I worked as a Research Fellow in the University of Social Welfare and Rehabilitation Sciences, Genetic Research Centre, and as a Senior Research Scientist in the Shahid Beheshti University of Medical Sciences, Division of Molecular and Cellular Endocrinology. During my time there, I wrote a research grant in the Genetic Research Centre titled as below:

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‐         Effective collaboration between NMDAR/Calpain, Wnt/Ca2+  and Wnt/β-catenin pathways is essential in normal development and function of human brain and regulation of CAPN10 gene expression.

 

I also participated in writing of a review paper “Diversity of Mutations in RET proto-oncogene and its mechanism of participation in Medullary Thyroid Cancer”, which was published in the "Journal of Critical Reviews in Clinical Laboratory of Science". In addition, I refereed a grant proposal in epigenetic in thyroid cancer and I was a reviewer for number of thyroid cancer related papers. I also supervised number of Medical Genetics PhD Students in the Genetic Centre.

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Previous to that, I worked in Dr Chris Elliott’s and Professor Alex Wade’s lab in collaboration with Lundbeck Pharmaceutical Company in Denmark. During my work there, which was published in Human Molecular Genetics (Afsari et al, 2014), I found that the visual gain control occurred faster in the one day old flies with G2019S mutation than the wild type fly. From these observations one could conclude that neuronal depolarization and repolarisation occurred differently in the mutated animals than the wild type. This fit with data published in (Hindle et al 2013) demonstrating that the process of pumping cataions across the plasma membrane  during dopaminergic  neurons depolarization  and  subsequent repolarization, required  extra  ATP produced by mitochondria. Therefore, increase in visual gain control in G2019S flies could well be coupled with high ATP demands and mitochondrial dysfunction,  which was highly recorded in the patients with Parkinson’s disease.

 

This  study  was  followed  by  investigating  the role  of number  of  LRRK2  kinase  inhibitors on  the  G2019S neuronal  gain control and examining whether any of these inhibitors  would rescue the neuronal gain control of G2019S mutant back to the wild type at the intermodulation level. (Inhibitors were LRRK2-IN-1  and  a novel  LRRK2 kinase inhibitor  BMPPB-32).

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This study demonstrated that both LRRK2-IN-1 and kinase inhibitor BMPPB-32 rescued the neuronal response of G2019S flies back to wild type. However, the specificity of these kinase inhibitors and their offtarget effects was examined using dLRRK- flies (without hLRRK2 and Drosophila own LRRK gene). After thorough examination of the effect of these kinase inhibitors on LRRK null flies, we concluded that BMPPB-32 was more specific as it did not have any offtarget effects; however, the LRRK2-IN-1 had effect on other kinases than the LRRK2 itself.

 

Previous to the aforementioned study, I worked on a project in Dr Paul Genever’s lab at University of York; Characterization of hTERT MSCs (human Telomerase Reverse Transcriptase Mesenchymal Stem Cells) and examined whether they differentiate to adipogenic and osteogenic lineages.

 

The clinical application of the mesenchymal stem cells as a resource for differentiation to adipogenic, osteogenic and chondrogenic lineages is a useful tool for tissue transplantation. However, there is a major obstacle with MSCs, as they have the capacity to become scenscence at the early stages of their growth. However, replicative ability of them can be increased  by immortalisation, and MSCs transduction by hTERT is one of the methods of achieving this goal.

 

Following above mentioned method of immortalisation, then these cells were grown exponentially in the culture for more than 400 days and their telomerase activity was confirmed by TRAPEZE  assay. Furthermore, their BMSC marker profile was identified by flow cytometry, which revealed positive  expression  for CD29, CD44, CD73, CD90, CD105 and CD166 and negative expression for CD34 and CD45 surface markers.

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The fact that Y101, Y102 and Y201 hTERT MSCs all possed spontaneous osteogenic differentiation capacity (immediately after being 100% confluent without addition of Dexamethason) made them a very good model for studying the osteogenesis. This was further facilitated as they did not readily differentiate to the adipogenic lineage even at the stage of 100% confluency; (exposure to adipogenic differentiating supplements is necessary for manifestation of this process).

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I successfully finished my first postdoctoral research fellowship at the Cardiovascular Centre of the University of Cincinnati, Ohio, USA in 2009. During those two years, I investigated the role of the canonical Wnt pathway in ischemic heart disease, which led to the finding of a novel mechanism that prevents apoptosis in hypoxic cardiomyocytes and therefore is cardioprotective. The paper was submitted in Cardiovascular Toxicology. In this paper I demonstrated that Wnt-3a induced the activation of canonical Wnt pathway through a PI3K/AKT dependent  pathway  in hypoxia.  HL-1 cardiomyocytes  were  left under  normoxic  or hypoxic condition for 24 hours. The activity of canonical Wnt pathway was demonstrated by immunoprecipitation  of β- catenin  and then immunoblotting for GSK3β  as  a  measure of the extent of  GSK3β-β-catenin complex formation. In this work, it was shown that at the basal level GSK3β-β catenin complexes were formed, meaning that there was little or no endogenous Wnt signalling activity. However, when the cells were pre-treated with recombinant Wnt-3a  protein (100 ng/ml),  the formation  of  GSK3β-β catenin  complex during hypoxia  was inhibited,  supporting that  there was an increased Wnt signalling.  Induction  of the canonical Wnt pathway activation in hypoxia was further confirmed using the TOP-Flash assay. TOP-flash assay demonstrated that in the cells co- transfected with Top-flash luciferase reporter plasmid, Wnt-3a treatment (100 ng/ml), resulted in an increase in transcriptional activation of TCF responsive genes, which further demonstrated the activation of canonical Wnt pathway in hypoxic cardiomyocyte.

 

PKB is activated through survival pathway when it is phosphorylated at Ser473. In HL-1 cardiomyocytes the level of phosphorylated PKB increased in Wnt-3a treated cells during hypoxia compared with the cells under normoxic condition.   This  suggested that  Wnt might activate PKB, which contributed to increased cell survival in Wnt-3a treated cardiomyocytes during hypoxia. In order to investigate this phenomenon, the HL-1 cardiomyocytes were transfected with PKB siRNA and treated with Wnt-3a under hypoxic condition, which led to a decrease in the level of BCL2 expression.  On the other hand, in the presence of just Wnt-3a in hypoxia the level of the BCL-2 increased, and further knock-down of PKB lowered the BCL-2 expression level.  Thus,  it was concluded that Wnt-3a activated the survival pathway in hypoxic cardiomyocytes through PKB dependent pathway.

 

My PhD focused on investigating the role of the Wnt signalling pathway in the mechanotransduction  pathway in SV-40 immortalised  human chondrocytes.  In this work it was demonstrated for the first time that induction of glycogen synthase kinase 3β (GSK3β)  activity  following  mechanical

stimulation  is  mediated  by  a  PI3K dependent pathway rather than a Wnt pathway. This was shown by a kinase assay development for GSK3β activity. In addition, the presence of GSK3-catenin complexes following mechanical stimulation and at the basal level in the presence and absence of Lithium (a Wnt agonist) were examined in two separate sets of

experiments.  These demonstrated that the formation of GSK3-catenin complex were induced after 5 minutes in the Lithium treated samples at the basal level and mechanical stimulation delayed this process (i.e complex formed after 10 min).

 

In addition, I demonstrated that two major heparan sulphate rich proteoglycan that  are highly expressed in cartilage, CD44 and fibroncetin, might act as a coreceptor for the Wnt-Fz ligand- receptor, in order to transduce the Wnt signal intracellularly.

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As an undergraduate  I chose to pursue a 4 year degree with a year research placement because I was eager to experience research at first hand as soon as possible.

 

During this placement I worked on determining microfilament and microtubule organization and dynamics  in CG-4 cells. These were done in order to establish CG-4 cells similarity to oligodendrocytes,  and  thus  demonstrating  the viability  of these cell  as  a  model  for exploring oligodendrocyte dynamics. This work led to two publications, Rumsby et al1998 and 2003.

 

During both my doctoral and post-doctoral research I have gained invaluable experience in mentoring students who contributed successfully to their field of research under my tutelage (7 undergraduates  who got 1st and 2:1 class degrees  and  2  technicians).  I believe  that  my supervision  and  technical  expertise in Drosophila neuroscience, Biochemistry, Molecular Cell  Biology,  Cell  Signalling, Physiology,  Genetics and  Stem  Cell  field  of  studies  makes  me an excellent Faculty for biological field of research