Affiliate Researchers

Annual Activity Report, March 2017

Prof. Farid Nakhoul - Single-nucleotide polymorphism (SNPs) in the autophagy-related gene 5 (ATG5) in patients with Diabetic Nephropathy & Retinopathy

The current study's aim is to evaluate whether or not a specific polymorphism (functionally silent difference) in the ATG5 gene is associated with diabetic renal and retinal diseases. Any day now, we should be receiving the final certification from the Helsinki Committee (medical research ethics committee) for this study. We are also reviewing the various DNA extraction kits, to find the optimal kit for our needs, so we can start calibrating our experimental system.

Prof. Jamal Zidan - Comparison of expression of breast cancer biomarkers in primary biopsies and in surgically resected tumors following neoadjuvant chemotherapy

Breast cancer patients usually undergo a core-needle biopsy (CNB) to determine the course of treatment. If the biopsy reveals the expression of certain biomarkers (namely, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor (HER2), and Ki67), then neoadjuvant chemotherapy (NAC) is indicated prior to surgical removal of the tumor. According to the medical literature the expression levels of these detective factors can change after NAC. We set out to measure these changes in our study, and the results may affect the choice of chemotherapy regimen the patient is given after surgery.

Dr. Andrei Braester, MD - Structural and Functional Characterization of Complement Components in Chronic Lymphocytic Leukemia Patients undergoing Immunotherapy

Dr. Andrei Braester, MD, Director, Institute of Hematology, Galilee Medical Center
Co-Investigator: Regina Michelis, PhD.
Researcher, Institute of Hematology, Galilee Medical Center

Chronic lymphocytic leukemia (CLL) is the most common adult leukemia in the western world. For treatment, clones of genetically designed white blood cells are used to trigger the body’s complementary immune system to target the cancer. Such an approach is called “complement-mediated cytotoxicity” (CDC). We wanted to study the structure of circulating complementary immune complexes and document the importance of their structural integrity for triggering an immune system attack during immunotherapy in patients with CLL.
Preliminary results indicate differences in the structure of complement complexes C4 and C5 in more than 50% of CLL patients, as well higher basal activity levels (indicated by C5b-9) as compared to healthy control subjects (HC). We have also documented that patients treated with aggregated IgG showed lower CDC responses than those treated with Zymosan. This indicates a possible link between the activation potential of the complement system in CLL patients and structural alterations in the C5 complex. It also suggests that developing a marker to detect changes in the C5 complex may assist clinicians in refining and personalizing the immunotherapeutic approach, improving CDC and consequently the therapy results.

Dr. Armaly Zaher - Impact of Haptoglobin Genotype on Intravenous Iron Administration-induced Oxidative Stress and Inflammatory response in Patients with Chronic Kidney Disease (CKD)

Background: Anemia is a common problem in CKD patients. It is attributed to decreased erythropoietin (EPO) production, low iron stores, and accompanying chronic inflammation. Therapy includes infusions of recombinant EPO to help boost erythropoietin production and also iron replenishment. However, treatment produces side effects, namely oxidative stress and inflammation, which are especially high in certain patients. L-carnitine supplementation has been used to mitigate these side effects in long-term dialysis patients. We were interested in how such supplementation would benefit early stage CKD patients. Furthermore, some patients have a variant form of haptoglobin, which protects the body by binding to and neutralizing loose hemoglobin in the bloodstream, which can cause kidney damage. We wanted to document the effect of this genetic variant on oxidative stress in CKD treatment.
Results: We found that combined administration of IVIR and carnitine brought about increased hemoglobin (Hb) levels in those subjects relative to subjects who received IVIR alone. Also, IVIR alone induced oxidative and inflammatory responses, but patients who received carnitine did not exhibit these adverse effects. These results were most profound in patients with the Hp2-2 variant of haptoglobin.
Conclusion: Our findings indicate that co-administration of carnitine with IVIR preferentially attenuates the adverse consequences of IVIR and suggests a role for carnitine therapy in these patients. Moreover, this study demonstrates that Hp2-2 is a significant risk factor for IVIR-induced oxidative stress in CKD patients.

Dr. Khaled Khazim and Dr. Cohen Idan - Genome-wide transcriptome profiling of circulating neutrophils in patients with end stage renal disease

In this research proposal we aimed to try and understand the underlying mechanism(s) of circulating blood neutrophils activation in ESRD patients using a full transcriptome analysis profiling. Our major goal was to try and uncover cellular mechanisms that will potentially lead to a better understanding of this process and may discover novel therapeutic targets for treatment or intervention. Additionally, by doing so, we hoped to discover a single "factor" of these cells that may be later used as a "biomarker" for assessing and monitoring atherosclerosis initiation and progression. We are happy to report that the mutual efforts of Dr. Idan Cohen, Din Richtert (the MD project student from the Faculty of Medicine) and I have already yielded 2-3 strong factors that play a pivotal role in neutrophils biology and can be used as potential biomarkers, including Hypoxia-inducible factor 1-alpha (HIF-1a), and Sirtuin 1 (SIRT-1).

Dr. Maya Wolf, MD - Fetal and maternal hormonal and signaling molecule association with outcome of labor induction

This study is a comparative evaluation of the correlation of fetal and maternal labor-associated hormones and signaling molecules between two methods of induction of labor in women with previous cesarean section. Collaborators are: Maya Wolf, MD, Department of Obstetrics & Gynecology, Galilee Medical Center, and the Faculty of Medicine, Bar-Ilan University, Israel; Jacob Bornstein, MD, MPA, Professor (PI) Department of Obstetrics & Gynecology, Galilee Medical center, and the Faculty of Medicine, Bar-Ilan University, Israel; Dr. Eilam Palzur – Research lab, Galilee Medical Center, Nahariya, Israel; Dr. Mona Shehada – the biochemistry lab, Galilee Medical Center, Nahariya, Israel
To date, 42 women have joined the study. The study aims include 60 women. The women were randomly divided into two groups: those who undergo induction of labor by balloon catheter or those who undergo breast stimulation. Blood samples for the analysis of hormone and signaling molecule levels were drawn at time 0, 3 and 6 hours from commencement of labor induction. In addition, immediately after delivery, cord blood was tested for those hormone and signaling molecules levels, and for bilirubin and cord pH. All samples are marked and preserved for later testing. Enrollment is expected to reach completion in two months, at which time all the samples will be processed and analyzed together, to ensure uniform laboratory conditions.

Prof. Offer Amir - miRNA in Acute Coronary Event: pathogenesis and risk stratification

Below is a brief progress report on our study of miRNA and its role in acute coronary events. Our study is aimed at providing ways of using miRNA to predict the onset of cardiac arrest and to identify an individual’s risk level for experiencing such an event.
We have optimized and adjusted the protocol for isolating blood components to our experimental settings, and performed a small pilot study. More than 300 patients have been recruited to the study so far. Blood samples were collected and processed. miRNA from 72 platelets samples was isolated and prepared for NGS. miRNA were sequenced on the HiSeq Illumina machine by Genomic Unit in the Faculty of Medicine, Safed. Bioinformatics analysis following the NGS was performed. Several miRNA were identified as differentially expressed. We intend to validate our findings using larger patient cohorts and a different experimental technique. We have chosen to use quantitative Real Time PCR (qPCR). So far we have optimized all steps of this procedure and adjusted for our experimental settings. To identify stable molecules suitable for our study, a NormFinder program was applied, with our NGS data used as an initial input.

Dr. Hadassah Goldberg & Dr. Rona Fell - Isolation and characterization of potentially metastasizing tumor cells from primary breast cancer samples

This is an interim report concerning a study funded by the Dangoor Foundation, 2016-17. The study is aimed at isolating breast cancer cells that harbor the potential to form distant metastases from primary tumors.
Our study is based on fresh tumor samples, which must be processed immediately after the surgical removal of the tumor. Therefore the entire study platform must be operative prior to initiation of sample collection. Up to now, we have completed the following steps:
1) We have constructed the method for tissue separation into a single cell suspension. 2) We have acquired the means necessary for removing blood stem cells from the total cell mixture. 3) We have conducted a comprehensive survey of the literature and selected the antibodies for separating the cell suspensions into two groups: metastasizing tumor cells (identified by markers of the ‘epithelial to mesenchymal transition’ (EMT) process), and non-metastasizing cells. This will be done using the Beckman Coulter MoFlo Astrios located at the Faculty of Medicine, Bar-Ilan University. 4) We have ordered the antibodies for removing stem cells. Once they arrive, we will start collecting tumor samples and begin the study. 5) We are grateful for the cooperation of Breast surgeon Dr. Assi Drobot and Pathologist expert Dr. Liat Appel from the Galilee Medical Center. Dr. Drobot has also agreed to present the study to the patients and ask them to sign an informed consent prior to their surgery.

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