Elucidation of Focal Adhesion Kinase as a Modulator of Migration and Invasion and as a Potential Therapeutic Target in Chronic Lymphocytic Leukemia

Thomas A. Burley; Andrew Hesketh; Giselda Bucca; Emma Kennedy; Eleni Ladikou; Benjamin Paul Towler; Simon Mitchell; Colin P. Smith; Christopher Fegan; Rosalynd Johnston; Andrea Pepper; Chris Pepper

doi:10.3390/cancers14071600

Elucidation of Focal Adhesion Kinase as a Modulator of Migration and Invasion and as a Potential Therapeutic Target in Chronic Lymphocytic Leukemia

Thomas A. Burley, Andrew Hesketh, Giselda Bucca, Emma Kennedy, Eleni Ladikou, Benjamin Paul Towler, Simon Mitchell, Colin P. Smith, Christopher Fegan, Rosalynd Johnston, Andrea Pepper, Chris Pepper

Research output: Contribution to journal › Article › peer-review

Abstract

The retention and re-migration of Chronic Lymphocytic Leukemia cells into cytoprotective and proliferative lymphoid niches is thought to contribute to the development of resistance, leading to subsequent disease relapse. The aim of this study was to elucidate the molecular processes that govern CLL cell migration to elicit a more complete inhibition of tumor cell migration. We compared the phenotypic and transcriptional changes induced in CLL cells using two distinct models designed to recapitulate the peripheral circulation, CLL cell migration across an endothelial barrier, and the lymph node interaction between CLL cells and activated T cells. Initially, CLL cells were co-cultured with CD40L-expressing fibroblasts and exhibited an activated B-cell phenotype, and their transcriptional signatures demonstrated the upregulation of pro-survival and anti-apoptotic genes and overrepresentation of the NF-κB signaling pathway. Using our dynamic circulating model, we were able to study the transcriptomics and miRNomics associated with CLL migration. More than 3000 genes were altered when CLL cells underwent transendothelial migration, with an overrepresentation of adhesion and cell migration gene sets. From this analysis, an upregulation of the FAK signaling pathway was observed. Importantly, PTK2 (FAK) gene expression was significantly upregulated in migrating CLL cells (PTK2 Fold-change = 4.9). Here we demonstrate that TLR9 agonism increased levels of p-FAK (p ≤ 0.05), which could be prevented by pharmacological inhibition of FAK with defactinib (p ≤ 0.01). Furthermore, a reduction in CLL cell migration and invasion was observed when FAK was inhibited (p ≤ 0.0001), supporting a role for FAK in both CLL migration and tissue invasion. When taken together, our data highlights the potential for combining FAK inhibition with current targeted therapies as a more effective treatment regime for CLL.

Original language	English
Article number	1600
Number of pages	16
Journal	cancers
Volume	14
Issue number	7
DOIs	https://doi.org/10.3390/cancers14071600
Publication status	Published - 22 Mar 2022

Bibliographical note

This work was supported by a Blood Cancer UK Programme Continuity Grant (18005), an MRC project grant (MR/V009095/1), the Sussex Cancer Fund, Crawley Leukaemia Research, and the Michael Chowen Research Fund, all awarded to A.G.S.P. and C.P., S.M. is funded by a Leukaemia UK John Goldman Fellowship (2020/JGF/003) and a UKRI Future Leaders Fellowship (MR/T041889/1).

Keywords

chronic lymphocytic leukemia
FAK
TLR9
migration
transcriptomics
miRNomics
microenvironment

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Burley, T. A., Hesketh, A., Bucca, G., Kennedy, E., Ladikou, E., Towler, B. P., Mitchell, S., Smith, C. P., Fegan, C., Johnston, R., Pepper, A., & Pepper, C. (2022). Elucidation of Focal Adhesion Kinase as a Modulator of Migration and Invasion and as a Potential Therapeutic Target in Chronic Lymphocytic Leukemia. cancers, 14(7), Article 1600. https://doi.org/10.3390/cancers14071600

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title = "Elucidation of Focal Adhesion Kinase as a Modulator of Migration and Invasion and as a Potential Therapeutic Target in Chronic Lymphocytic Leukemia",

abstract = "The retention and re-migration of Chronic Lymphocytic Leukemia cells into cytoprotective and proliferative lymphoid niches is thought to contribute to the development of resistance, leading to subsequent disease relapse. The aim of this study was to elucidate the molecular processes that govern CLL cell migration to elicit a more complete inhibition of tumor cell migration. We compared the phenotypic and transcriptional changes induced in CLL cells using two distinct models designed to recapitulate the peripheral circulation, CLL cell migration across an endothelial barrier, and the lymph node interaction between CLL cells and activated T cells. Initially, CLL cells were co-cultured with CD40L-expressing fibroblasts and exhibited an activated B-cell phenotype, and their transcriptional signatures demonstrated the upregulation of pro-survival and anti-apoptotic genes and overrepresentation of the NF-κB signaling pathway. Using our dynamic circulating model, we were able to study the transcriptomics and miRNomics associated with CLL migration. More than 3000 genes were altered when CLL cells underwent transendothelial migration, with an overrepresentation of adhesion and cell migration gene sets. From this analysis, an upregulation of the FAK signaling pathway was observed. Importantly, PTK2 (FAK) gene expression was significantly upregulated in migrating CLL cells (PTK2 Fold-change = 4.9). Here we demonstrate that TLR9 agonism increased levels of p-FAK (p ≤ 0.05), which could be prevented by pharmacological inhibition of FAK with defactinib (p ≤ 0.01). Furthermore, a reduction in CLL cell migration and invasion was observed when FAK was inhibited (p ≤ 0.0001), supporting a role for FAK in both CLL migration and tissue invasion. When taken together, our data highlights the potential for combining FAK inhibition with current targeted therapies as a more effective treatment regime for CLL.",

keywords = "chronic lymphocytic leukemia, FAK, TLR9, migration, transcriptomics, miRNomics, microenvironment",

author = "Burley, {Thomas A.} and Andrew Hesketh and Giselda Bucca and Emma Kennedy and Eleni Ladikou and Towler, {Benjamin Paul} and Simon Mitchell and Smith, {Colin P.} and Christopher Fegan and Rosalynd Johnston and Andrea Pepper and Chris Pepper",

note = "This work was supported by a Blood Cancer UK Programme Continuity Grant (18005), an MRC project grant (MR/V009095/1), the Sussex Cancer Fund, Crawley Leukaemia Research, and the Michael Chowen Research Fund, all awarded to A.G.S.P. and C.P., S.M. is funded by a Leukaemia UK John Goldman Fellowship (2020/JGF/003) and a UKRI Future Leaders Fellowship (MR/T041889/1).",

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TY - JOUR

T1 - Elucidation of Focal Adhesion Kinase as a Modulator of Migration and Invasion and as a Potential Therapeutic Target in Chronic Lymphocytic Leukemia

AU - Burley, Thomas A.

AU - Hesketh, Andrew

AU - Bucca, Giselda

AU - Kennedy, Emma

AU - Ladikou, Eleni

AU - Towler, Benjamin Paul

AU - Mitchell, Simon

AU - Smith, Colin P.

AU - Fegan, Christopher

AU - Johnston, Rosalynd

AU - Pepper, Andrea

AU - Pepper, Chris

N1 - This work was supported by a Blood Cancer UK Programme Continuity Grant (18005), an MRC project grant (MR/V009095/1), the Sussex Cancer Fund, Crawley Leukaemia Research, and the Michael Chowen Research Fund, all awarded to A.G.S.P. and C.P., S.M. is funded by a Leukaemia UK John Goldman Fellowship (2020/JGF/003) and a UKRI Future Leaders Fellowship (MR/T041889/1).

PY - 2022/3/22

Y1 - 2022/3/22

N2 - The retention and re-migration of Chronic Lymphocytic Leukemia cells into cytoprotective and proliferative lymphoid niches is thought to contribute to the development of resistance, leading to subsequent disease relapse. The aim of this study was to elucidate the molecular processes that govern CLL cell migration to elicit a more complete inhibition of tumor cell migration. We compared the phenotypic and transcriptional changes induced in CLL cells using two distinct models designed to recapitulate the peripheral circulation, CLL cell migration across an endothelial barrier, and the lymph node interaction between CLL cells and activated T cells. Initially, CLL cells were co-cultured with CD40L-expressing fibroblasts and exhibited an activated B-cell phenotype, and their transcriptional signatures demonstrated the upregulation of pro-survival and anti-apoptotic genes and overrepresentation of the NF-κB signaling pathway. Using our dynamic circulating model, we were able to study the transcriptomics and miRNomics associated with CLL migration. More than 3000 genes were altered when CLL cells underwent transendothelial migration, with an overrepresentation of adhesion and cell migration gene sets. From this analysis, an upregulation of the FAK signaling pathway was observed. Importantly, PTK2 (FAK) gene expression was significantly upregulated in migrating CLL cells (PTK2 Fold-change = 4.9). Here we demonstrate that TLR9 agonism increased levels of p-FAK (p ≤ 0.05), which could be prevented by pharmacological inhibition of FAK with defactinib (p ≤ 0.01). Furthermore, a reduction in CLL cell migration and invasion was observed when FAK was inhibited (p ≤ 0.0001), supporting a role for FAK in both CLL migration and tissue invasion. When taken together, our data highlights the potential for combining FAK inhibition with current targeted therapies as a more effective treatment regime for CLL.

AB - The retention and re-migration of Chronic Lymphocytic Leukemia cells into cytoprotective and proliferative lymphoid niches is thought to contribute to the development of resistance, leading to subsequent disease relapse. The aim of this study was to elucidate the molecular processes that govern CLL cell migration to elicit a more complete inhibition of tumor cell migration. We compared the phenotypic and transcriptional changes induced in CLL cells using two distinct models designed to recapitulate the peripheral circulation, CLL cell migration across an endothelial barrier, and the lymph node interaction between CLL cells and activated T cells. Initially, CLL cells were co-cultured with CD40L-expressing fibroblasts and exhibited an activated B-cell phenotype, and their transcriptional signatures demonstrated the upregulation of pro-survival and anti-apoptotic genes and overrepresentation of the NF-κB signaling pathway. Using our dynamic circulating model, we were able to study the transcriptomics and miRNomics associated with CLL migration. More than 3000 genes were altered when CLL cells underwent transendothelial migration, with an overrepresentation of adhesion and cell migration gene sets. From this analysis, an upregulation of the FAK signaling pathway was observed. Importantly, PTK2 (FAK) gene expression was significantly upregulated in migrating CLL cells (PTK2 Fold-change = 4.9). Here we demonstrate that TLR9 agonism increased levels of p-FAK (p ≤ 0.05), which could be prevented by pharmacological inhibition of FAK with defactinib (p ≤ 0.01). Furthermore, a reduction in CLL cell migration and invasion was observed when FAK was inhibited (p ≤ 0.0001), supporting a role for FAK in both CLL migration and tissue invasion. When taken together, our data highlights the potential for combining FAK inhibition with current targeted therapies as a more effective treatment regime for CLL.

KW - chronic lymphocytic leukemia

KW - FAK

KW - TLR9

KW - migration

KW - transcriptomics

KW - miRNomics

KW - microenvironment

U2 - 10.3390/cancers14071600

DO - 10.3390/cancers14071600

M3 - Article

VL - 14

JO - cancers

JF - cancers

IS - 7

M1 - 1600

ER -

Elucidation of Focal Adhesion Kinase as a Modulator of Migration and Invasion and as a Potential Therapeutic Target in Chronic Lymphocytic Leukemia

Abstract

Bibliographical note

Keywords

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