Life Sciences & Biotechnology
Title : | Development of chemically modified aptamers for targeting neuronal exosomes |
Area of research : | Life Sciences & Biotechnology |
Focus area : | Neuroscience |
Principal Investigator : | Dr. Manish Debnath, CSIR-Indian Institute Of Chemical Biology (CSIR–IICB), West Bengal |
Timeline Start Year : | 2024 |
Timeline End Year : | 2026 |
Contact info : | manish.debnath@iicb.res.in |
Details
Executive Summary : | Exosomes are membrane-bound extracellular vesicles of ~140 nm size that can encapsulate important intracellular biomolecules such as proteins, nucleic acids, and lipids and transport them to the surrounding environment. These exosomes play an important role in the development of brain disorders like Parkinson's disease (PD), which is the second most important neurodegenerative disease. Unfortunately, there are very few strategies that allow the early detection of PD and even fewer therapeutic opportunities are available. PD is caused by the presence of lewy bodies and clumps formed by a protein called α-synuclein. At the onset of PD, the diseased neuronal cells pack the α-synuclein aggregates within exosomes and secrete them in the biofluid. When these exosomes in biofluids fuse with other neuronal cells, the aggregated diseased form of α-synuclein may spread and accumulate in different parts of the brain, leading to the development of PD. Since PD exosomes are critical for cell-cell communication and disease development, they may serve as important targets to combat PD. Moreover, neuronal exosomes of the diseased phenotype (e.g. PD) are pathologically different from the aparkinsonian exosomes and could serve as a biomarker to diagnose PD at a very early (prodromal) stage. In this project, we would synthesize chemically modified RNA aptamers (synthetic antibodies, CemRmers) that could identify and target the PD exosomes. These CemRmers will be synthesized using Exo-SELEX in which a large library of RNA is screened against PD-exosomes and the selected RNA molecules are amplified enriched in a multi-step process. The chemical modifications will be introduced into the selected aptamers by an in-vitro transcription method, which will lead to the synthesis of CemRmers with high specificity and physiological stability. The CemRmers will be biorthogonally linked to PEGylated magnetic nanoparticles to further improve their biocompatibility and bioavailability. Overall, the proposed work has four prime objectives: 1) To isolate and validate the neuronal exosomes relevant to PD. 2) To synthesise CemRmers against PD exosomes utilizing the Exo-SELEX approach; 3) To develop CemRmer-nanoparticle conjugates for point-of-care diagnosis of PD by employing hybridization chain reaction; 4) To investigate the ability of CemRmer-nanoparticle conjugates to capture and neutralize the exosomes in biofluids. The design of these CemRmer-nanoparticle conjugates against exosomes is highly original. The strategy to target exosomes for combating PD and the use of HCR as a diagnostic tool are also less explored. The proposed work is interdisciplinary in nature and will utilize my past expertise in nucleic acid research, molecular biology, and membrane biophysics to solve a problem in neurobiology. Development of this system may provide quick detection of PD from patient samples and may pave the way for future development of therapeutics against PD. |
Total Budget (INR): | 27,76,160 |
Organizations involved