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The role of Bcl-2 proteins in regulating metabolism in normal and tumour cells

Lead Research Organisation: Babraham Institute
Department Name: UNLISTED

Abstract

Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.

Technical Summary

Tumour cells exhibit a number of properties that set them apart from normal cells. Understanding what makes a tumour cell different from a normal cell is important as it may allow us to identify new ways to selectively kill tumour cells. Tumour cells differ markedly from normal cells in the way they generate energy, being far more dependent upon the metabolism of glucose (a process known as glycolysis) than normal cells. We are interested in determining if this is an ‘Achilles heel’ by which we can attack tumour cells. If we disrupt glycolysis do tumour cells die? If so, how do they die? If not then, are they now more vulnerable to death in response to other drugs?

Planned Impact

unavailable
 
Description We have demonstrated links between the abundance of NAD+, a critical cofactor in hundreds of metabolic reactions, and the BCL2 protein family. Specifically reduction in the abundance of NAD+ (which os observed durign ageing) results in the activation of BCL2 celld eath pathway.
Exploitation Route We are currently exploring links between NAD+ abundance, autophagy and cell death pathways. This may be relevant to normal ageing but also to certian diseases that are more prevalent as we age. We will explore this through discussiosn with the Biotech/Pharma sector
Sectors Education

Healthcare

Pharmaceuticals and Medical Biotechnology

 
Description Impacts form this work are still emerging but we anticipate the results will be of interest to the Pharma sector and may provide new insights for treating disease
Sector Healthcare,Pharmaceuticals and Medical Biotechnology
Impact Types Economic

 
Title BIM KO, BMF KO and BIM/BMF DKO CRISPR cell lines - A375 cells 
Description A375 lines with deletions of BIM, BMF or both BIM and BMF generated by CRISPR/Cas9 and validated 
Type Of Material Cell line 
Year Produced 2020 
Provided To Others? Yes  
Impact These cell lines have been used to define the mechanism of action of a novel drug combination that effectively kills melanoma cells Targeting melanoma's MCL1 bias unleashes the apoptotic potential of BRAF and ERK1/2 pathway inhibitors. Sale MJ, Minihane E, Monks NR, Gilley R, Richards FM, Schifferli KP, Andersen CL, Davies EJ, Vicente MA, Ozono E, Markovets A, Dry JR, Drew L, Flemington V, Proia T, Jodrell DI, Smith PD, Cook SJ. Nat Commun. 2019 Nov 14;10(1):5167. doi: 10.1038/s41467-019-12409-w 
URL https://pubmed.ncbi.nlm.nih.gov/31727888/
 
Title BIM KO, BMF KO and BIM/BMF DKO CRISPR cell lines - SW620 cells 
Description SW620 cell lines with deletions of BIM, BMF or both BIM and BMF generated by CRISPR/Cas9 and validated 
Type Of Material Cell line 
Year Produced 2020 
Provided To Others? No  
Impact These cells have been used to define the mechanism of action of a novel drug combination that effectively kills colorectal cancer cells 
 
Title BIM/BMF DKO CRISPR cell lines - SK-MEL-30 
Description SK-MEL-30 lines with deletions of both BIM and BMF generated by CRISPR/Cas9 and validated 
Type Of Material Cell line 
Year Produced 2020 
Provided To Others? Yes  
Impact These cell lines have been used to define the mechanism of action of a novel drug combination that effectively kills melanoma cells Targeting melanoma's MCL1 bias unleashes the apoptotic potential of BRAF and ERK1/2 pathway inhibitors. Sale MJ, Minihane E, Monks NR, Gilley R, Richards FM, Schifferli KP, Andersen CL, Davies EJ, Vicente MA, Ozono E, Markovets A, Dry JR, Drew L, Flemington V, Proia T, Jodrell DI, Smith PD, Cook SJ. Nat Commun. 2019 Nov 14;10(1):5167. doi: 10.1038/s41467-019-12409-w 
URL https://pubmed.ncbi.nlm.nih.gov/31727888/
 
Title Mammalian expression plasmids for BIM mutants 
Description Mammalian expression plasmids encoding a variety of splice variants and mutants of the pro-apoptotic protein BIM, including phospho-site mutants 
Type Of Material Technology assay or reagent 
Provided To Others? Yes  
Impact New insights into the post-translational regulation of the pro-apoptptic protein BIM. 
 
Description AMPK signalling 
Organisation Imperial College London
Department Institute of Clinical Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution We have been investigating the role of the AMPK kinase in detecting changes in cellular NAD+ levels
Collaborator Contribution Provision of a small molecule agonist of AMPK
Impact None yet but a manuscript is in preparation
Start Year 2015
 
Description AstraZeneca ERK 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution We want to understand the various mechanisms by which ERK1/2 pathway-addicted tumour cells adapt and acquire resistance to the MEK1/2 inhibtor Selumetinib. We have generated a variety of human tumour cell lines with acquired resistance to the clinical candidate MEK1/2 inhibitor Selumetinib. These include cell lines in which BRAF is the driving oncogene but also those in which KRAS is the driving oncogene. We have analysed the activation state of the ERK1/2 pathway in these cells and in some cases validated the resistance mechanism. In other cases this analysis is ongoing. In some models we find that resistance is reversible upon drug withdrawal suggesting that resistant cells actually have a fitness deficit in the absence of drug. We are investigating the mechanisms that underlie this fitness deficit. For our partner (AstraZeneca/CRUK) this may allow development of rational strategies to overcome or delay resistance and thereby provide more durable drug responses. For our own basic biological interests this should provide insights into how this key cell fate signalling pathway is regulated; this may be relevant to emerging regenerative medicine protocols.
Collaborator Contribution AstraZeneca have performed a variety of Next Gen Seq analyses on samples provided by us to identify genetic and transcriptomic changes associated with resistance; these may be candidate resistance drivers. They are also performing xenograft studies to test specific hypotheses that emerge form our studies. Our collaborators at the University of Bath are investigating how the magnitude of ERK1/2 signalling can impart different tumour cell responses when Selumetinib is withdrawn from resistant cells. In a three-way collaboration between our lab, the CRUK-CI and AstraZeneca we are peforming high throughput drug combination screens to identify drugs that combine with Selumetinib to provide superior tumour growth inhibition or tumour cell death.
Impact Much of this is still early stage. However, high throughput drug screening has identified several drug combinations that markedly transform the growth inhibitory effects of Selumetinib. Our analysis of new resistance models has identified several completely novel potential mechanisms which we are in the process of validating. These studies will lead to further papers in addition those publications already reported and may contribute to the testing of new drug combinations in the clinic in the future.
Start Year 2013
 
Description AstraZeneca ERK 
Organisation Cancer Research UK Cambridge Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution We want to understand the various mechanisms by which ERK1/2 pathway-addicted tumour cells adapt and acquire resistance to the MEK1/2 inhibtor Selumetinib. We have generated a variety of human tumour cell lines with acquired resistance to the clinical candidate MEK1/2 inhibitor Selumetinib. These include cell lines in which BRAF is the driving oncogene but also those in which KRAS is the driving oncogene. We have analysed the activation state of the ERK1/2 pathway in these cells and in some cases validated the resistance mechanism. In other cases this analysis is ongoing. In some models we find that resistance is reversible upon drug withdrawal suggesting that resistant cells actually have a fitness deficit in the absence of drug. We are investigating the mechanisms that underlie this fitness deficit. For our partner (AstraZeneca/CRUK) this may allow development of rational strategies to overcome or delay resistance and thereby provide more durable drug responses. For our own basic biological interests this should provide insights into how this key cell fate signalling pathway is regulated; this may be relevant to emerging regenerative medicine protocols.
Collaborator Contribution AstraZeneca have performed a variety of Next Gen Seq analyses on samples provided by us to identify genetic and transcriptomic changes associated with resistance; these may be candidate resistance drivers. They are also performing xenograft studies to test specific hypotheses that emerge form our studies. Our collaborators at the University of Bath are investigating how the magnitude of ERK1/2 signalling can impart different tumour cell responses when Selumetinib is withdrawn from resistant cells. In a three-way collaboration between our lab, the CRUK-CI and AstraZeneca we are peforming high throughput drug combination screens to identify drugs that combine with Selumetinib to provide superior tumour growth inhibition or tumour cell death.
Impact Much of this is still early stage. However, high throughput drug screening has identified several drug combinations that markedly transform the growth inhibitory effects of Selumetinib. Our analysis of new resistance models has identified several completely novel potential mechanisms which we are in the process of validating. These studies will lead to further papers in addition those publications already reported and may contribute to the testing of new drug combinations in the clinic in the future.
Start Year 2013
 
Description AstraZeneca ERK 
Organisation University of Bath
Country United Kingdom 
Sector Academic/University 
PI Contribution We want to understand the various mechanisms by which ERK1/2 pathway-addicted tumour cells adapt and acquire resistance to the MEK1/2 inhibtor Selumetinib. We have generated a variety of human tumour cell lines with acquired resistance to the clinical candidate MEK1/2 inhibitor Selumetinib. These include cell lines in which BRAF is the driving oncogene but also those in which KRAS is the driving oncogene. We have analysed the activation state of the ERK1/2 pathway in these cells and in some cases validated the resistance mechanism. In other cases this analysis is ongoing. In some models we find that resistance is reversible upon drug withdrawal suggesting that resistant cells actually have a fitness deficit in the absence of drug. We are investigating the mechanisms that underlie this fitness deficit. For our partner (AstraZeneca/CRUK) this may allow development of rational strategies to overcome or delay resistance and thereby provide more durable drug responses. For our own basic biological interests this should provide insights into how this key cell fate signalling pathway is regulated; this may be relevant to emerging regenerative medicine protocols.
Collaborator Contribution AstraZeneca have performed a variety of Next Gen Seq analyses on samples provided by us to identify genetic and transcriptomic changes associated with resistance; these may be candidate resistance drivers. They are also performing xenograft studies to test specific hypotheses that emerge form our studies. Our collaborators at the University of Bath are investigating how the magnitude of ERK1/2 signalling can impart different tumour cell responses when Selumetinib is withdrawn from resistant cells. In a three-way collaboration between our lab, the CRUK-CI and AstraZeneca we are peforming high throughput drug combination screens to identify drugs that combine with Selumetinib to provide superior tumour growth inhibition or tumour cell death.
Impact Much of this is still early stage. However, high throughput drug screening has identified several drug combinations that markedly transform the growth inhibitory effects of Selumetinib. Our analysis of new resistance models has identified several completely novel potential mechanisms which we are in the process of validating. These studies will lead to further papers in addition those publications already reported and may contribute to the testing of new drug combinations in the clinic in the future.
Start Year 2013
 
Description Lecture in Cancer Biology and Medicine training course 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Postgraduate students
Results and Impact I presented a lecture on how signalling pathways are remodelled to drive innate or acquired resistance to new targeted anti-cancer agents that are clincially approved or in development. The lecturer reached approx 60 Master and PhD students, clinicians some patient advocates and charity-funded researchers. There was a vibrant follow-up Q&A session and new contacts were established with the prospect of future collaborations. In feedback >80% of the audience found it useful
Year(s) Of Engagement Activity 2020,2021
 
Description Science Open Day 
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 Students visited the lab and undertook small lab-based proejcts supervised by students/post-docs and myself. I explained the research that we do and discussed ethical issues such as the use of animals in research.
This precipitated excellent discussion and dialogue.

We received excellent feedback from the schools involved and requests for further outreach activities
Year(s) Of Engagement Activity 2013,2014,2015,2016,2017,2018,2019,2020