Live Single Cell RNAseq on Treatment Resistant MCF7 Breast Cancer cells
Anthony Monteza Cabrejos a
a Institute of Chemical Biology, Molecular Sciences Research Hub, Imperial College London, 82 Wood Lane, London W12 0BZ, UK
b Department of Chemistry, Molecular Sciences Research Hub, Imperial College London, 82 Wood Lane, London W12 0BZ, UK
Proceedings of Emerging Investigators in Microfluidics Conference (EIMC)
Online, Spain, 2021 July 20th - 21st
Organizers: Adrian Nightingale, Darius Rackus and Claire Stanley
Poster, Anthony Monteza Cabrejos, 042
Publication date: 5th July 2021
ePoster: 

Breast cancer is the most prevalent cancer in women worldwide with oestrogen dependent breast cancer representing 70% of all new diagnosis1. There is still much left to learn from the individual genomic changes that occur during tumorigenesis and in response to treatment. Approximately ~3% of the patients each year return with overt relapse, inevitably leading to further metastatic development1. The frequency of relapse is constant for up to 20 years post-surgery1 making Endocrine Therapy resistance one the most important problems clinically for patient management. The processes of adaptation and selection leading to this late relapse are poorly understood and this is limited by current research tools available. In current science we know that tracking genomic changes is possible. Genomic changes are irreversible and permanently documented by the genome. Current, approaches to study cell genomics and transcriptomics involves the isolation and extraction of nucleic acids from individual cells and the generation of next-generation sequencing libraries. The drawback with this method is that the sample is destroyed each time. A central question in treatment resistant breast cancer is to understand how the cells evolve during early oncogenesis and treatments. Currently, the ability to track dynamic transcriptional changes is unachievable, as each time point requires the exhaustion of the sample. We aim to develop a step changing technology for the next generation of high throughput minimally invasive nanotweezers for high throughout RNA sampling and sequencing whilst keeping the cell alive and viable. The nanotweezers consist of two closely spaced electrodes with gaps as small as 10-20nm, which can be used for dielectrophoretic targeted trapping of nucleic acids, proteins, and even single organelles2. They can be spatially controlled to extract from living cells with single molecule precision. This technique does not aspirate cytoplasmic fluid and allows for pre-concentration of analyte in real time, making it better than AFM and Nanopipettes2. These nanotweezers have further been improved in their ability to trap mRNA in cells through the addition of Poly Thiamine oligomer sequence at the tip (functionalised). These functionalised nanotweezers are subsequently used to extract and monitor mRNA levels of previously identified genes involved in the eventual development of treatment resistance.

Thank you to Dr. Binoy Paulose Nadappuram, Dr. Sung Pil Hong, and Debjani Saha for their support and guidance throughout my project. 

Thank you to Professor Edel, Professor Ivanov and Professor Magnani for supporting my ideas in the is project and facilitating this research. 

Lastly, thank you to EPSRC for funding me to do this project.

© Fundació Scito
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