Metabolomics applications to in-cell NMR based processes for monitoring the uptake of drugs and biologics

Lead Research Organisation: University of Cambridge
Department Name: Biochemistry

Abstract

The development portfolio is changing as many therapeutic targets are not "drugable" with conventional approaches using either small molecules or antibody proteins. Nucleic acid based therapeutics offer the possibility to engage with these targets, but only if the major translational challenges can be addressed. Optimal intracellular delivery is the bottleneck for the advancement of nucleic-acid based therapies. However, a central question is posed with the development of many new treatments. Have we got what we want, where we want it?

Foremost amongst the challenges is selecting and developing effective and safe transfection vectors for in-vivo studies. A major gap at present is that our existing in-vitro models are poorly predictive of animal studies. We will use NMR spectroscopy based methods to help bridge this gap in this studentship. Current in-vitro approaches to determine effective cellular uptake typically make extensive use of microscopy based methods using labelled nucleic acids. Here we propose to use High Resolution Magic Angle Spinning (HRMAS) NMR spectroscopy to measure the metabolic response of whole living cells to treatment. The cells would then be available for conventional analysis too. By multivariate statistical analysis it is possible to use this technique to:

Determine the metabolic profile of living cells prior to and following treatment with anti-cancer drugs.
Infer cellular uptake based on metabolic profile.
Propose a mechanism of action based on changes in the cell's metabolic profile.

In this studentship we will use a combination of HRMAS 1H NMR spectroscopy and mass spectrometry based approaches to model the uptake and effects of biologics and drugs in a range of cell lines to develop an in vitro test for drug uptake at the start of the development process. Should this project be successful it will contribute significantly to reducing the number of animals used in drug toxicity studies by providing a simple and robust in vitro test for the uptake of compounds into cells.

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
EP/P510440/1 01/10/2016 30/09/2021
1789837 Studentship EP/P510440/1 01/10/2016 30/09/2020 Marta Wylot
 
Description I am exploring possible applications of high resolution magic angle spinning NMR to monitor changes in intact cells induced by apoptotic agents.
So far I have measured diffusion rates of lipid droplets in intact cells and observed significant changes between control and apoptotic cells.
Main findings:
1. Lipid droplet diffusion rates in intact cells measured by HRMAS NMR for the first time correlates well with microscopy results and literature values of diffusion rates measured in vivo by MRS.
2. Diffusion measurements in conjunction with 1H NMR-derived lipid signal provide a novel means of following apoptosis.
Exploitation Route My findings may help to understand lipid metabolism during apoptosis; possible application in drug development process.
(early stage, project in progress)
Sectors Pharmaceuticals and Medical Biotechnology

 
Description 11th NMR: a tool for biology Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact A number of conference participants stopped by to see my poster and hear about my projects. A couple of PhD student were interested in applying similar methods for their projects. We exchanged contacts and are in touch. I also had an interesting discussion about ideas how I can extend my current project and the person was happy to get involved.
Year(s) Of Engagement Activity 2019
URL http://www.nmr2019.conferences-pasteur.org
 
Description Cambridge Science Festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact General public visited our stand to watch our engaging demonstrations and learn interesting facts. Children were keen to participate in our activities and asked a lot of questions. We received many positive comments from parents about the activity.
Year(s) Of Engagement Activity 2017
URL https://www.sciencefestival.cam.ac.uk
 
Description EBI-Sanger-Cambridge PhD Symposium 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact The symposium was organized for PhD students to share their research and scientific interests. I had discussions with a number of PhD students about my research and exchanged contact details with two people who were interested in my project.
Year(s) Of Engagement Activity 2019
URL https://escamps.org/escamps19
 
Description King's College Natural Sciences Taster session 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Gave a mini lecture to sixth formers interested in coming to Cambridge University. Held a question and answers session after the lecture and discussed the teaching at the University.
Year(s) Of Engagement Activity 2018
 
Description Metabolomic conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact A number of conference participants stopped by to see my poster and hear about my projects. Three PhD students worked on projects using the same technique and we had interesting discussions and exchanged troubleshooting tips and protocols for analysing samples and data.
Year(s) Of Engagement Activity 2018
URL http://metabolomics2018.org
 
Description Pint of Science 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact 30 people attended 3 talks about science in past present and future. After each talk there were a number of questions from audience. We organized a hands on activate for younger audience involving decorating safety secs and we promoted active learning through a pub quiz. After an event participants engaged in scientific discussions.
Year(s) Of Engagement Activity 2017
URL https://pintofscience.co.uk
 
Description The Naked Scientist Podcast 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Talked about my research while in a punt going along the river Cam as part of the Naked scientist Podcast.
Year(s) Of Engagement Activity 2018