Gene Expression Profiling of FFPE Samples: A Titration Test.

The gene expression analysis of formalin-fixed paraffin-embedded (FFPE) tissues is often hampered by poor RNA quality, which results from the oxidation, cross-linking and other chemical modifications induced by the inclusion in paraffin. Yet, FFPE samples are a valuable source for molecular studies and can provide great insights into disease progression and prognosis. With the advancement of genomic technologies, new methods have been established that offer reliable and accurate gene expression workflows on samples of poor quality. NanoString is a probe-based technology that allows the direct counting of the mRNA transcripts and can be applied to degraded samples. Here, we have tested 2 RNA extraction methods for FFPE samples, and we have performed a titration experiment to evaluate the impact of RNA degradation and RNA input on the gene expression profiles assessed using the NanoString IO360 panel. We have selected FFPE samples of different DV200 values and assessed them on the nCounter platform with 2 different amounts of input RNA. This study concludes that the nCounter is a robust and reliable platform to assess the gene expression of RNA samples with DV200 > 30%; its robustness and ease of use could be of particular benefit to clinical settings.

Enhancing Mesenchymal Stromal Cell Immunomodulation for Treating Conditions Influenced by the Immune System.

Mesenchymal stromal cells (MSCs), formerly known as mesenchymal stem cells, are nonhematopoietic multipotent cells and are emerging worldwide as the most clinically used and promising source for allogeneic cell therapy. MSCs, initially obtained from bone marrow, can be derived from several other tissues, such as adipose tissue, placenta, and umbilical cord. Diversity in tissue sourcing and manufacturing procedures has significant effects on MSC products. However, in 2006, a minimal set of standard criteria has been issued by the International Society of Cellular Therapy for defining derived MSCs. These include adherence to plastic in conventional culture conditions, particular phenotype, and multilineage differentiation capacity in vitro. Moreover, MSCs have trophic capabilities, a high in vitro self-renewal ability, and immunomodulatory characteristics. Thus, immunosuppressive treatment with MSCs has been proposed as a potential therapeutic alternative for conditions in which the immune system cells influence outcomes, such as inflammatory and autoimmune diseases. The precise mechanism by which MSCs affect functions of most immune effector cells is not completely understood but involves direct contact with immune cells, soluble mediators, and local microenvironmental factors. Recently, it has been shown that their homeostatic resting state requires activation, which can be achieved in vitro with various cytokines, including interferon-γ. In the present review, we focus on the suppressive effect that MSCs exert on the immune system and highlight the significance of in vitro preconditioning and its use in preclinical studies. We discuss the clinical aspects of using MSCs as an immunomodulatory treatment. Finally, we comment on the risk of interfering with the immune system in regard to cancer formation and development.

Tailoring cells for clinical needs: Meeting report from the Advanced Therapy in Healthcare symposium (October 28-29 2017, Doha, Qatar).

New technologies and therapies designed to facilitate development of personalized treatments are rapidly emerging in the field of biomedicine. Strikingly, the goal of personalized medicine refined the concept of therapy by developing cell-based therapies, the so-called "living drugs". Breakthrough advancements were achieved in this regard in the fields of gene therapy, cell therapy, tissue-engineered products and advanced therapeutic techniques. The Advanced Therapies in Healthcare symposium, organized by the Clinical Research Center Department of Sidra Medicine, in Doha, Qatar (October 2017), brought together world-renowned experts from the fields of oncology, hematology, immunology, inflammation, autoimmune disorders, and stem cells to offer a comprehensive picture of the status of worldwide advanced therapies in both pre-clinical and clinical development, providing insights to the research phase, clinical data and regulatory aspects of these therapies. Highlights of the meeting are provided in this meeting report.

Harnessing the immune system for the treatment of melanoma: current status and future prospects.

When malignant melanoma is diagnosed early, surgical resection is the intervention of choice and is often curative, but many patients present with unresectable disease at later stages. Due to its complex etiology paired with well-documented chemoresistance and high metastatic potential, patients with advanced melanoma had a poor prognosis, and the treatment of this disease remained unsatisfactory for many years. Recently, targeted therapy, immune checkpoint inhibition, or combinatory approaches have revolutionized the therapeutic options of melanoma allowing considerable improvement in disease control and survival. In this review we will summarize these novel therapeutic strategies with particular focus on combinatory immunotherapies and further discuss recent data derived from immunogenomic studies and potential options to improve the therapeutic efficacy of immune modulatory approaches.

Risk factors contributing to road traffic crashes in a fast-developing country: the neglected health problem.

BACKGROUND: The main objective of this study was to explore the patterns of road traffic crashes (RTCs) among drivers in Qatar and to examine the contributing factors involved. METHODS: This study was a cross-sectional survey conducted from February to June 2009 at the Primary Health Care Centers in the State of Qatar. A random sample of 1675 Qatari drivers were approached, and 1228 of them responded and agreed to participate in this study, with a response rate of 74.6%. Face to face interview was conducted by well-trained research assistants based on a questionnaire covering sociodemographic information, driving history, type of vehicle, driver behavior, details of crashes, and accident pattern. RESULTS: The study revealed that 26.6% of the studied Qatari drivers were involved in RTCs. 69.4% of the drivers with a history of RTCs were male. Drivers in the age group 25-34 years showed a higher risk of having a crash (31.2%). Drivers of cars (45.6%) and 4WD/SUVs (23.5%) were more frequently involved in RTCs. 23.5% of drivers who had RTC did not always wear their seat belt while driving. 37.9% of the drivers with RTC were engaged in eating and drinking and 41.9% were using their mobile phones while driving. More than half of the drivers with RTCs had traffic violations (57.2%), with 25.7% exceeding the speed limit. A highly significant difference was observed between the two groups (drivers with and without RTC) in terms of presence of traffic violation (<0.001) and exceeding the speed limit (<0.001). Most of the crashes happened during sunny days (84.7%) with fewer crashes on holidays (5.5%) and weekends (12.5%). Overturn skid (17.7%), angle collision (14.3%) and rear-end hit (10.7%) were the most frequently reported patterns of RTCs. CONCLUSION: The study findings revealed the high risk of RTCs among drivers in the State of Qatar. Among the sociodemographic factors, male drivers and young drivers aged 25-34 years were found to have a higher accident involvement. Human behavior was identified as the main contributing factor of all RTCs, especially presence of traffic violations.