Rajgopal Srinivasan, Ph.D.
Principal Scientist & Head Bio IT R&D, TCS Innovation Labs, India
Interpretation of Genomic Variation – Identifying Rare Variations Leading to Disease
Genome sequencing technologies are generating an abundance of data on human genetic variations. A big challenge lies in interpreting the functional relevance of such variations, especially in clinical settings. A first step in understanding the clinical relevance of genetic variations is to annotate the variants for region of occurrence, degree of conservation both within and across species, pattern of variation across related individuals, novelty of the variation and know effects of related variations. Several tools already exist for this purpose. However, these tools have their strengths and weaknesses. A second issue is the development of algorithms, which, given a rich annotation of variants are able to prioritize the variants as being relevant to the phenotype under investigation.
In my talk I will detail work that has been done in our labs to address both of the above problems. I will also illustrate the application of these tools that helped identify a rare mutation in the ATM gene leading to a diagnosis of AT in two infants.
Manzoor K, Ph.D.
Professor, Centre for Nanoscience & Molecular Medicine, Amrita University
Targeting aberrant cancer kinome using rationally designed nano-polypharmaceutics
Manzoor Koyakutty, Archana Ratnakumary, Parwathy Chandran, Anusha Ashokan, and Shanti Nair
`War on Cancer’ was declared nearly 40 years ago. Since then, we made significant progress on fundamental understanding of cancer and developed novel therapeutics to deal with the most complex disease human race ever faced with. However, even today, cancer remains to be the unconquered `emperor of all maladies’. It is well accepted that meaningful progress in the fight against cancer is possible only with in-depth understanding on the molecular mechanisms that drives its swift and dynamic progression. During the last decade, emerging new technologies such as nanomedicine could offer refreshing life to the `war on cancer’ by way of providing novel methods for molecular diagnosis and therapy.
In the present talk, we discuss our approaches to target critically aberrant cancer kinases using rationally designed polymer-protein and protein-protein core-shell nanomedicines. We have used both genomic and proteomic approaches to identify many intimately cross-linked and complex aberrant protein kinases behind the drug resistance and uncontrolled proliferation of refractory leukemic cells derived from patients. Small molecule inhibitors targeted against oncogenic pathways in these cells were found ineffective due to the involvement of alternative survival pathways. This demands simultaneous inhibition more than one oncogenic kinases using poly-pharmaceutics approach. For this, we have rationally designed core-shell nanomedicines that can deliver several small molecules together for targeting multiple cancer signalling. We have also used combination of small molecules and siRNA for combined gene silencing together with protein kinase inhibition in refractory cancer cells. Optimized nanomedicines were successfully tested in patient samples and found enhanced cytotoxicity and molecular specificity in drug resistant cases.
Nano-polypharmaceutics represents a new generation of nanomedicines that can tackle multiple cancer mechanisms simultaneously. Considering the complexity of the disease, such therapeutic approaches are not simply an advantage, but indispensable.
Acknowledgements:
We thank Dept. of Biotechnology and Dept. Of Science and Technology,Govt. of India for the financial support through `Thematic unit of Excellence in Medical NanoBiotechnology’ and `Nanomedicine- RNAi programs’.
H S M Kort, J-W J Lammers, S N W Vorrink, T Troosters
Introduction
Chronic Obstructive Pulmonary Disease (COPD) is a disabling airway disease with variable extrapulmonary effects that may contribute to disease severity in individual patients (Rabe et al. 2007). The world health organization predicts that COPD will become the third leading cause of death worldwide by 2030. Patients with COPD demonstrate reduced levels of spontaneous daily physical activity (DPA) compared with healthy controls (Pitta et al. 2005). This results in a higher risk of hospital admission and shorter survival (Pitta et al. 2006). Pulmonary rehabilitation can help to improve the DPA level, however, obtained benefits decline after 1–2 years (Foglio et al. 2007).
Purpose
In order to maintain DPA in COPD patients after rehabilitation, we developed a mobile phone application. This application measures DPA as steps per day, measured by the accelerometer of the smartphone, and shows the information to the patient via the display of the mobile phone. A physiotherapist can monitor the patient via a secure website where DPA measurements are visible for all patients. Here, DPA goals can be adjusted and text messages sent.
Method
Three pilot studies were performed with healthy students and COPD patients to test the application for usability, user friendliness and reliability with questionnaires and focus groups. Subjects also wore a validated accelerometer. For the Randomized Controlled Trial (RCT) 140 COPD patients will be recruited in Dutch physiotherapy practises. They will be randomised in an intervention group that receives the smartphone for 6 months and a control group. Measurements include lungfunction, dyspnea, and exercise capacity and are held at 0, 3, 6 and 12 months.
Results and Discussion
The application was found to be useful, easy to learn and use. Subjects had no problems with health care professionals seeing information on their physical activity performance. They do find it important to be able to determine who can see the information. Correlations between the accelerometer and the measurements on DPA of the smartphone for steps per hour were 0.69 and 0.70 for pilot studies 1 (students) and 2 (COPD patients) respectively. The version of the application in pilot study 3 contained an error, which made correlations with the accelerometer unusable. The RCT study is now being executed.
Shrikant Anant, Ph.D.
The Department of Molecular & Integrative Physiology, Kansas University Medical Center, USA
Cancer Stem Cells: Target Colon Cancers
Shrikant Anant, Deep Kwatra and Dharmalingam Subramaniam
Colon cancer is a leading cause of cancer related deaths in the US, and its rate is increasing at an alarming rate in lndia. Recent studies have suggested the drug resistance role for a mall number of cells within a tumor called cancer stem cells. We identified the colon cancer stem cell marker DCLK1, a member of the protein kinase superfamily and the doublecortin family. The protein encodes a Cterminal serinethreonine protein kinase domain, which shows substantial homology to Ca2calmodulindependent protein kinase. Our current studies have been to identify compounds that can either affect DCLK1 expression or inhibits its activity as a way to inhibit cancer stem cells. Honokiol is a biphenolic compound that has been used in the traditional Chinese Medicine for treating various ailments. In vitro kinase assays with recombinant DCLK1 demonstrated that honokiol inhibits its kinase activity in a dose dependent manner. We therefore determined the effect of honokiol on stem cells. One method to look at effects on stem cells is perform a spheroid assay, where spheroids formation is suggested to maintain stemlike characteristic of cancer cells. Honokiol significantly suppressed colonosphere formation of two colon cancer cell lines HCT116 and SW480. Flow cytometry studies confirmed that honokiol reduced the number of DCLK1cells. A critical signaling pathway known to modulate intestinal stem cell proliferation is the Hippo signaling pathway, and deregulation of the pathway leads to tumor development. DCLK1cells had high levels of YAP1, the nuclear target of Hippo signaling. We determined the effect of honokiol on components of the hipposignaling pathway. Honokiol reduced the phosphorylation of Mst1/2, Lats1/2 and YAP1. Furthermore, honokiol treatment resulted in downregulation of YAPTEAD complex protein TEAD-1. Ectopic expression of the TEAD-1 partially rescued the cells from honokiol mediated growth suppression. To determine the effect of honokiol on tumor growth in vivo, nude mice harboring HCT116 tumor xenografts in their flanks were administered the compound intraperitoneally every day for 21 days. Honokiol treatment significantly inhibited tumor xenograft growth. Western blot and immunohistochemistry analyses demonstrated significant inhibition in the expression of stem marker and Hippo signaling proteins in the honokioltreated xenograft tissues. Taken together, these data suggest that honokiol is a potent inhibitor of colon cancer that targets DCLK1 stem cells by inhibiting Hippo signaling pathway.