Finding the sweet spot in the Matrix: allowing immune cell migration but not tumour dissemination

Primary supervisor: Alberto Elosegui-Artola, Francis Crick Institute

Secondary supervisor: Victoria Sanz-Moreno, Queen Mary University of London

Project

Immunotherapies are one of the most promising strategies against cancer. However, there is a lack of in vitro models to comprehend cancer and immune cells dynamics in response to these therapies. These in vitro models need to recapitulate the tumour microenvironment (TME), especially the extracellular matrix (ECM) properties. Even though immune cells must navigate the ECM, little is known about the influence of the ECM mechanical properties, such as viscoelasticity, in the density and diversity of tumour-infiltrating immune cells. The aim of this project is to develop a versatile in vitro model to determine the influence of the ECM mechanics in immune cell response versus cancer cell invasion and determine the best strategy to tackle cancer. The main aims are:

1. To understand, with a novel microfluidic system, the influence of the ECM viscoelasticity in immune cell migration/activation (CD8+ T cells and macrophages) versus cancer cell invasion.
2. To determine the molecular mechanism that regulates immune cell migration in response to ECM mechanical properties.
3. To perform a high-throughput drug screening to study the role of the ECM mechanical properties in therapy responses and validate these results in vivo.

Firstly, we will collaborate with the Francis Crick Institute’s Making Lab to develop a microfluidic-based in vitro model. To mimic the aberrant TME, the mechanical properties of the ECM in the device will be modified our recently developed viscoelastic hydrogel system (Elosegui-Artola et al. 2022). The microfluidic device design will allow us to perform high-spatiotemporal resolution microscope experiments. By combining parallel measurements of immune cell migration and tumour spheroids invasion and fate, we will determine the role of ECM mechanics in anti-tumour response. We hypothesize that the ECM mechanics will not only regulate immune cells’ migratory ability but also their activity. We will measure T cell check exhaustion markers (i.e. PD-1, LAG- 3, etc) and checkpoint molecules such as PDL1 in macrophages and cancer cells.

Secondly, we will elucidate the molecular mechanism that regulates ECM mechanics driven anti-tumour response. We have previously shown that the cellular force-generation machinery composed of integrins, adaptor proteins (i.e. talin) and the actomyosin cytoskeleton regulates how cells respond to ECM mechanics (Elosegui-Artola et al. 2014, 2016, 2017). Additionally, we found that non-muscle Myosin II supports not only cancer cell actomyosin driven migration, but also cancer cell survival and cancer cell-immune cell communication (Georgouli et al. 2019, Orgaz et al. 2020, Rodriguez-Hernandez et al. 2020). A cytoskeleton directed phosphoprotein array (Georgouli et al. 2019) will be used to elucidate which mechano-transducing molecules are regulated by ECM mechanics in cancer cells and in immune cells. We expect to find the mechanosensitive sweet spot that activates immune cell infiltration while preventing cancer cell invasion.

Lastly, the screening in aim 2 will reveal different potential kinases as regulators of anti-tumoral response. We will take advantage of our high-throughput system to screen a subset of small molecule inhibitors directed to kinases to determine the influence of mechanics in therapy. We will collaborate with the Francis Crick Isntitute’s High- throughput Screening facility to develop this aim. We will test our most promising drug candidate in vivo, where we will measure tumour growth, cytoskeletal phospho-changes and immune infiltration using multiplex digital pathology pipelines.

Candidate background

The candidate should be enthusiastic about understanding how the interplay between mechanical signals and biochemical signals regulates tumour response. This project would suit candidates with a background in bioengineering, biology, biophysics, biochemistry.

Potential Research Placements

1. Making Lab, Francis Crick Institute
2. Oliver Pearce, Barts Cancer Institute, Queen Mary University of London
3. Ilaria Malanchi, Francis Crick Institute

References

1. Elosegui-Artola A, Gupta A, Najibi A, Seo BR, Garry R, De Lázaro I, Tringides C, Darnell M, W Gu, Q Zhou, DA Weitz, L Mahadevan, DJ Mooney (2022) Matrix Viscoelasticity controls spatio-temporal tissue organization. Biorxiv. [link]
2. Orgaz JL, Crosas-Molist E, Sadok A, Perdrix-Rosell A, Maiques O, Monger J, Rodriguez-Hernandez I, Mele S, Georgouli M, Bridgeman V, Karagiannis P, Pandya P, Cantelli G, Boehme L, Wallberg F, V Tape C, Karagiannis SN, Malanchi I, Sanz-Moreno V (2020) Myosin II reactivation and Cytoskeletal remodelling as a hallmark and a vulnerability in melanoma resistance. Cancer Cell, 37, 85-103. [link]
3. Rodriquez-Hernandez I, Maiques O, Kohlhammer L, Cantelli G, Pedrix-Rosell A, Monger J, Fanshawe B, Bridgeman VL, Karagiannis S, Penin RM, Marcolval J, Marti RM, Matias-Guiu Z, Fruhwirth GO, Orgaz JL, Malanchi I, Sanz-Moreno V (2020) WNT11-FZD7-DAAM1 signalling supports tumour initiating abilities and melanoma amoeboid invasion. Nature Communications. [link]
4. Georgouli M, Herraiz C, Crosas-Molist E, Fanshawe B, Maiques O, Perdrix A, Pandya P, Cantelli G, Rodriguez-Hernandez I, Karagiannis P, Lam H, Josephs D, Matias-Guiu X, Marti RM, Nestle FO, Orgaz JL, Malanchi I, Fruhwirth GO, Karagiannis SN and Sanz-Moreno V (2019) Regional activation of Myosin II in cancer cells drives tumour progression via a secretory cross-talk with the immune microenvironment. Cell. [link]
5. Elosegui-Artola A, Oria R, Chen Y, Kosmalska K, Pérez-González C, Castro N, Zhu C, Trepat X, Roca- Cusachs P. (2016) Mechanical regulation of a molecular clutch defines force transmission and transduction in response to matrix rigidity. Nature Cell Biology. [link]