Vural Özdemir Ph.D.
Sanjeeva Srivastava Ph.D.
Krishnakumar Menon, Ph.D.
Associate Professor, Centre for Nanosciences & Molecular Medicine, Amrita University, Kochi, India
A Far-Western Clinical Proteomics Approach to Detect Molecules of Clinical and Pathological Significance in the Neurodegenerative Disease Multiple Sclerosis.
Multiple Sclerosis (MS), an autoimmune neurodegenerative disorder of the central nervous system. The disease affects young adults at their prime age leading to severe debilitation over several years. Despite advances in MS research, the cause of the disease remains elusive. Thus, our objective is to identify novel molecules of pathological and diagnostic significance important in the understanding, early diagnosis and treatment of MS. Biological fluids such as cerebrospinal fluid (CSF), that bathe the brain serve as a potential source for the identification of pathologically significant autoantibody reactivity in MS. In this regard, we report the development of an unbiased clinical proteomics approach for the detection of reactive CSF molecules that target brain proteins from patients with MS. Proteins of myelin and myelin-axolemmal complexes were separated by two-dimensional gel electrophoresis, blotted onto membranes and probed separately with biotinylated unprocessed CSF samples. Protein spots that reacted specifically to MS-CSF’s were further analyzed by matrix assisted laser desorption ionization-time-of-flight time-of-flight mass spectrometry. In addition to previously reported proteins found in MS, we have identified several additional molecules involved in mitochondrial and energy metabolism, myelin gene expression and/or cytoskeletal organization. Among these identified molecules, the cellular expression pattern of collapsin response mediator protein-2 and ubiquitin carboxy-terminal hydrolase L1 were investigated in human chronic-active MS lesions by immunohistochemistry. The observation that in multiple sclerosis lesions phosphorylated collapsin response mediator protein-2 was increased, whereas Ubiquitin carboxy-terminal hydrolase L1 was down-regulated, not only highlights the importance of these molecules in the pathology of this disease, but also illustrates the use of our approach in attempting to decipher the complex pathological processes leading to multiple sclerosis and other neurodegenerative diseases. Further, we show that in clinicaly isolated syndrome (CIS), we could identify important molecules that could serve as an early diagnostic biomarker in MS.
Claudia AM Wheeler-Kingshott, Ph.D.
University Reader in Magnetic Resonance Physics, Department of Neuroinflammation, UCL Institute of Neurology, London, UK
Abstract
Detecting neuronal activity in vivo non-invasively is possible with a number of techniques. Amongst these, in 1990 functional magnetic resonance imaging (fMRI) was proposed as a technique that has a great ability to spatially map brain activity by exploiting the blood oxygenation level dependent (BOLD) contrast mechanism [1, 2]. In fact, neuronal activation triggers a demand for oxygen and induces a localised increase in blood flow and blood volume, which actually exceeds the metabolic needs. This in turns causes an increase of oxyhaemoglobin in the venous compartment, which is a transient phenomenon and is accompanied by a transient change (decrease) in the concentration of deoxyhaemoglobin. Due to its paramagnetic properties, the amount of deoxyhaemoglobin present in the venous blood affects the local magnetic field seen by the spins (protons) and determines the local properties of the MR signal. A decrease in deoxyhaemoglobin during neuronal activity, therefore, induces local variations of this magnetic field that increases the average transverse relaxation time of tissue, measured via the T2* parameter [3]. This means that there is an increase of the MR signal (of the order of a few %, typically <5%) linked to metabolic changes happening during brain function. Activation can be inferred at different brain locations by performing tasks while acquiring the MR signal and comparing periods of rest to periods of activity.
The macroscopic changes of the BOLD signal are well characterised, while the reason for the increased blood supply, exceeding demands, needs further thoughts. Here we will discuss two approaches for explaining the BOLD phenomenon, one that links it to adenosine triphosphate production [4] and enzyme saturation, the other that relates it to the very slow diffusion of oxygen through the blood-brain-barrier with a consequent compensatory high demand of oxygen [5]. Some evidence of restricted oxygen diffusion has been shown by means of hypercapnia [6], although it is not excluded that both mechanisms may be present.
Overall, the BOLD signal changes theory and its physiological basis will be presented and discussed.
References
- Ogawa, S., et al., Brain magnetic resonance imaging with contrast dependent on blood oxygenation. Proc Natl Acad Sci U S A, 1990. 87(24): p. 9868-72.
- Kwong, K.K., et al., Dynamic magnetic resonance imaging of human brain activity during primary sensory stimulation. Proc Natl Acad Sci U S A, 1992. 89(12): p. 5675-9.
- Bandettini PA, et al. Spin-echo and gradient-echo EPI of human brain activation using BOLD contrast: a comparative study at 1.5 T. NMR Biomed. 1994 Mar;7(1-2):12-20
- Fox, P.T., et al., Nonoxidative glucose consumption during focal physiologic neural activity. Science, 1988. 241(4864): p. 462-4.
- Gjedde, A., et al. Reduction of functional capillary density in human brain after stroke. J Cereb Blood Flow Metab, 1990. 10(3): p. 317-26.
- Hoge, R.D., et al., Linear coupling between cerebral blood flow and oxygen consumption in activated human cortex. Proc Natl Acad Sci U S A, 1999. 96(16): p. 9403-8.
Vural Özdemir, MD, Ph.D., DABCP
Co-Founder, DELSA Global, Seattle, WA, USA
Crowd-Funded Micro-Grants to Link Biotechnology and “Big Data” R&D to Life Sciences Innovation in India
Vural Özdemir, MD, PhD, DABCP1,2*
- Data-Enabled Life Sciences Alliance International (DELSA Global), Seattle, WA 98101, USA;
- Faculty of Management and Medicine, McGill University, Canada;
ABSTRACT
Aims: This presentation proposes two innovative funding solutions for linking biotechnology and “Big Data” R&D in India with artisan small scale discovery science, and ultimately, with knowledge-based innovation:
- crowd-funded micro-grants, and
- citizen philanthropy
These two concepts are new, and inter-related, and can be game changing to achieve the vision of biotechnology innovation in India, and help bridge local innovation with global science.
Background and Context: Biomedical science in the 21(st) century is embedded in, and draws from, a digital commons and “Big Data” created by high-throughput Omics technologies such as genomics. Classic Edisonian metaphors of science and scientists (i.e., “the lone genius” or other narrow definitions of expertise) are ill equipped to harness the vast promises of the 21(st) century digital commons. Moreover, in medicine and life sciences, experts often under-appreciate the important contributions made by citizen scholars and lead users of innovations to design innovative products and co-create new knowledge. We believe there are a large number of users waiting to be mobilized so as to engage with Big Data as citizen scientists-only if some funding were available. Yet many of these scholars may not meet the meta-criteria used to judge expertise, such as a track record in obtaining large research grants or a traditional academic curriculum vitae. This presentation will describe a novel idea and action framework: micro-grants, each worth $1000, for genomics and Big Data. Though a relatively small amount at first glance, this far exceeds the annual income of the “bottom one billion” – the 1.4 billion people living below the extreme poverty level defined by the World Bank ($1.25/day).
We will present two types of micro-grants. Type 1 micro-grants can be awarded through established funding agencies and philanthropies that create micro-granting programs to fund a broad and highly diverse array of small artisan labs and citizen scholars to connect genomics and Big Data with new models of discovery such as open user innovation. Type 2 micro-grants can be funded by existing or new science observatories and citizen think tanks through crowd-funding mechanisms described herein. Type 2 micro-grants would also facilitate global health diplomacy by co-creating crowd-funded micro-granting programs across nation-states in regions facing political and financial instability, while sharing similar disease burdens, therapeutics, and diagnostic needs. We report the creation of ten Type 2 micro-grants for citizen science and artisan labs to be administered by the nonprofit Data-Enabled Life Sciences Alliance International (DELSA Global, Seattle: http://www.delsaglobal.org). Our hope is that these micro-grants will spur novel forms of disruptive innovation and life sciences translation by artisan scientists and citizen scholars alike.
Address Correspondence to:
Vural Özdemir, MD, PhD, DABCP
Senior Scholar and Associate Professor
Faculty of Management and Medicine, McGill University
1001 Sherbrooke Street West
Montreal, Canada H3A 1G5
Email: vural.ozdemir@alumni.utoronto.ca
