Tspan6 as a regulator of immune microenvironment in breast cancer

Lead Research Organisation: University of Birmingham
Department Name: Cancer Sciences


Growth of tumour is controlled at the multiple levels which involved changes in tumour cell responses to extracellular signals. Conversely, tumour cells themselves modify surrounding tissue by reorganising its architecture and cellular composition. It is very important to understand how tumour cells cross communicate with microenvironment as this will help to design novel therapeutic approaches.

The proposed work is focussed on breast cancer and specifically centred on examining new pathways of communications between tumour cells and B-cells which contribute to anti-tumour host defence. We have discovered that the abundance of a cell surface molecule called Tspan6 is decreased during cancerous progression. Importantly, loss or decrease in the level of Tspan6 correlated with worse patient survival. We have also noticed that when Tspan6 was abundant on breast tumour cells the cancerous tissues were enriched in a specific type of lymphocytes. This observation led us to hypothesise that when expressed in breast cancer cells Tspan6 might control the process anti-tumour host responses. The outlined programme will investigate molecular processes that would explain this phenomenon. Specifically, we will use breast cancer tissues to examine whether Tspan6 exhibits specific preferences towards various subtypes of lymphocytes in directing their ability to accumulate in cancerous tissues. We will identify Tspan6-dependent molecular factors derived from tumour cells that control accumulation of lymphocytes in cancerous tissues. We will also examine how Tspan6 change an intracellular network of signalling processes that underly the ability of tumour cells to influence recruitment and/or retention of B-cells in cancerous tissues.

Technical Summary

The immune cell landscape within tumour microenvironment is controlled by an elaborate network of communications involving chemokines, cytokines and growth factors which are produced by the immune cells themselves, by cancer cells and by other cell types in cancerous tissues (e.g. tumour endothelium, tumour-associated fibroblasts). Whilst specific contribution of various types of tumour-infiltrating T-cells and macrophages to tumour development and host-immune responses is becoming clearer, significantly less is known and understood about the function of tumour-infiltrating B-cells (TIL-B).

Whilst studying the immune landscape in breast cancer we discovered that the abundance of TIL-B correlated with the expression of tetraspanin 6 (Tspan6) by tumour cells. Importantly, Tspan6-expressing breast cancer cells stimulated migration of B-lymphocytes in vitro. These observations point towards a novel pathway underlying communication between tumour cells and TIL-B. As the expression of Tspan6 positively correlated with patients survival, we hypothesise that that Tspan6 suppresses breast cancer progression by modulating tumour infiltration of B-cells and, consequently, the immune landscape of tumour tissues. The proposed work aims to investigate the molecular pathways underlying the phenomenon of increased B-cell chemotaxis towards Tspan6-expressing breast cancer cells.
We will:
-establish the contribution of chemotactic factors differentially produced by Tspan6-expressing breast cancer cells.
-delineate Tspan6-dependent pathways in breast cancer cells that control infiltration of B-cells to cancerous tissues.
-examine the B-cell profile in breast cancer tissues in relation to expression of Tspan6.
-evaluate the role of Tspan6-dependent recruitment of B-cells to cancerous tissues in vivo.

Planned Impact

The applicants are already actively involved in outreach activities which are organised by the School of Cancer Sciences in Birmingham. This involves lectures to the general public and lab tours. All have excellent contacts with clinical colleagues at the University Hospital Birmingham and Heartlands Hospital. The principal applicant has recently organised an International Symposium on the Inflammatory breast cancer which involved patient advocates and was covered in a press release by the Breast Cancer Now.

All these activities will continue during the life-time of the grant.

Commercial engagement:
Pharmaceutical and biotech companies are learning about the technologies being developed here through partnerships and visits to Birmingham. The applicants will take further steps to engage with pharma and biotech companies.
Description The role of tetraspanin CD151 in inflammatory breast cancer
Amount £23,780 (GBP)
Organisation The Inflammatory Breast Cancer Network UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2019 
End 06/2019
Description The role of tetraspanin proteins in the development of inflammatory breast cancer
Amount £37,000 (GBP)
Organisation The Inflammatory Breast Cancer Network UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2018 
End 12/2018
Description tetraspanin proteins in IBC
Amount £157,633 (GBP)
Organisation University of Birmingham 
Sector Academic/University
Country United Kingdom
Start 01/2021 
End 06/2021
Title Tspan6 knockout animals 
Description Tspan6 knockout animals on the FVB genetic background 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2018 
Provided To Others? No  
Impact Generation of this strain of knockout animals will be useful as a model system for breast cancer research. 
Title cultures of breast cancer organoid 
Description We have started routinely generating culture of breast cancer organoids from cancerous and normal tissues. These culture will allow us to test the effect of current and newly develop therapeutics and represent a more physiological platform for drug testing. 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? No  
Impact We are currently testing these models in a number of projects including those aimed at investigation of the tetraspanin-dependent regulation of the immune microenvironment. 
Title IBC cohort 
Description We have collated a clinical dataset of the inflammatory breast cancer patients which are currently using to analyse the data on the immune microenvironment in IBC. 
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? No  
Impact We will be able to examine the IBC immune microenvironment in detail for the first time. 
Description Tetraspanins and extracellular vesicles 
Organisation University Medical Center Freiburg
Country Germany 
Sector Hospitals 
PI Contribution We hypothesised that tetraspanin protein Tspan6 regulates production and composition of the extracellular vesicles (EVs). We send the material for the analysis.
Collaborator Contribution Our collaborators are using the nanoparticle-tracking assay to examine the concentration and size distribution of EVs produced by breast cancer cells
Impact The manuscript is currently in preparation.
Start Year 2019
Description Tetraspanins and oxysterol production 
Organisation Aston University
Department School of Life and Health Sciences
Country United Kingdom 
Sector Academic/University 
PI Contribution We discovered that extracellular vesicles (EVs) produced by breast cancer cells stimulate migration of B cells in oxysterol-dependent manner. We purified EVs from the culture supernatant generated by breast cancer cells.
Collaborator Contribution Quantitative analysis of EV-associated oxysterols by mass spectrometry.
Impact Manuscript is currently in preparation...
Start Year 2019
Description analysis of Tspan6 expression in clinical samples 
Organisation Palacky University
Department Institute of Molecular and Translational Medicine
Country Czech Republic 
Sector Academic/University 
PI Contribution We have collected a cohort of clinical samples for this research project
Collaborator Contribution partners performed immunohistochemical staining and primary statistical analysis of the data.
Impact The work has ben presented at the European tetraspanin conference in 2019. Generated data that guide our research project.
Start Year 2016
Description bioinformatics support 
Organisation Antwerp University Hospital
Country Belgium 
Sector Hospitals 
PI Contribution We have outlined the aims of the projects and provided data
Collaborator Contribution Our collaborator, Professor S.van Laere performed bioinformatics analysis.
Impact this collaboration has already led to one joint publication with further two manuscripts in their final stages of preparation
Start Year 2019
Description examination of the immune cell dynamics in breast cancer in vivo 
Organisation University of Birmingham
Department School of Immunity and Infection
Country United Kingdom 
Sector Academic/University 
PI Contribution We provide mouse cell model to study the role of tetraspanin proteins in the regulation of the immune microenvironment
Collaborator Contribution Our collaborator provides a syngeneic mouse animal model to study the role of tetraspanin proteins in the regulation of the immune microenvironment
Impact Not yet.
Start Year 2019
Description Interview to the BBC Spanish language service 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact answered specific questions posed by a reporter
Year(s) Of Engagement Activity 2017
Description Member of the Sceintific advisory board at the Breast cancer Now 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Member of the Scientific advisory board at the Breast Cancer Now
Year(s) Of Engagement Activity 2014,2015,2016,2017,2018,2019