Studying Single Cell Protein Signalling Dynamics using High-throughput Microfluidic Platform to Elucidate the role of Inflammation in Neuropathology.


CALL: 2013

DOMAIN: BM - Regenerative Medicine in Age-related Diseases

FIRST NAME: Suresh Kumar

LAST NAME: Poovathingal



HOST INSTITUTION: University of Luxembourg

KEYWORDS: Single cell ProteomicsMicrofluidicsInflammationNeuro-PathologyProtien Signalling NetworkComputational Biology

START: 2013-05-01

END: 2014-04-30


Submitted Abstract

Role of inflammation in neuropathology has long been appreciated. However, the molecular mechanisms of initiation and subsequent progression of the pathology remains largely elusive, predominantly due to the complexity of interactions of different cellular macromolecules in cells. Comprehending these mechanisms amidst the underlying complexities requires quantification of cellular biomolecules at single-cell level using high-throughput “omics” assays and development of complementary modelling techniques to characterize the resulting high dimensional data. One of the prime objectives of this secondment is to develop such an integrative platform for specifically studying the role of inflammation in neuropathology/degeneration. During inflammation there exists a diverse array of molecular cross talks in different neuronal cell types, and an intricate network of these biochemical elements controls the inflammation process and keeps the process under close check. To understand the role of inflammation in neurodegeneration and neuro-pathology, it is important to learn how the inflammatory responses are induced and propagated in the context of different molecular and cellular interactions and the mechanism by which these responses ultimately contribute to pathology. Different neuronal cell-types such as neurons, microglia and neuro- epithelial or endothelial cell will be considered in this work. Common inflammation mediators such as TNF-a and IFN-¿ will be used for perturbations. Cellular systems are finite and the biochemical interactions existing in the cellular processes are discrete, stochastic in nature. A plethora of evidence indicate cellular heterogeneity commonly exists within an isogenic or clonal population of cells. Conventional bulk cell-based quantification assays measure the average response from a large population of cells, with an assumption that average response is representative of typical cell attributes. However, this is not true in many cases, and the measurements obtained from bulk-assays are often masked by the average response from majority of cells. This would render the signals from rare cells in the population to get averaged out, which in several cases are more pertinent to the investigation (a good example for this would be certain drug resistant cancer cells, which are responsible for tumour relapse after therapy). Resolving such stochastic dynamics of biomolecular interactions necessitates single-cell-level analysis and to derive statistically meaningful data by performing analysis over several single cells. Thus, it becomes critical to develop high throughput platform, which has the capability of single-cell quantification. Among different techniques capable of performing single-cell-level quantification, most promising technologies capable of producing high throughput biological data are: single-cell immunostaining, and flow cytometry methods. However, aside from the presentation of non-physiological conditions to the cells, these techniques are also quite cumbersome. In recent years, microfluidics has emerged as a powerful enabling technology that has immense potential for capturing the inherent complexity of cellular systems by providing high throughput biological data with higher accuracy from limited number of cells. Also, the fabrication and handling of this technology is relatively simpler. Hence, the primary objective of this visit is to develop microfluidic based proteomic assays for single-cell-level quantification and to gain expertise and knowledge transfer (to Luxembourg) in emerging fields of micro-fabrication and microfluidics. Also, in the due course set up a long-term working collaboration with pioneers in microfluidics technology.

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