p53 family members and apoptosis in malignant melanoma: a role in chemoresistance?
Lead Research Organisation:
Queen Mary, University of London
Department Name: Blizard Institute of Cell and Molecular
Abstract
We are investigating why melanoma, an aggressive form of skin cancer, is so resistant to chemotherapy, and how new treatments might be able to overcome this resistance. In the UK 8,100 new cases of melanoma are diagnosed annually with the incidence predicted to treble in the next 30 years. Melanoma arises from the pigment-producing cells (melanocytes) in skin and both its development and resistance to standard cancer treatments is thought in part due to the resistance of cells to apoptosis - a form of cell death. Essential to apoptosis is the p53 gene and loss of p53 activity is frequently implicated in other cancers. p53 also plays a crucial role in protecting against skin cancer induced by ultraviolet radiation (UVR). The aim of this project is to study the p53 family of proteins and their interacting roles in UVR-induced cell death in melanocytes and melanoma. Melanoma represents a major burden on public health and healthcare resources. This project will determine the mechanism of p53 inactivation in melanoma. Information obtained will further our understanding of changes in melanocytes as they undergo malignant progression and provide new ways to develop treatments that are effective.
Technical Summary
Malignant melanoma is the most aggressive form of skin cancer. Its incidence has increased more rapidly than that of any other cancer in the last 30 years. Advanced disease has poor prognosis and is notoriously resistant to all current therapeutic modalities. Unlike many other malignancies, p53 mutations are infrequent in melanoma implicating aberrant upstream or downstream components of the p53 signalling pathway. In the proposed study the hypothesis that the p53 family members are determinants of chemosensitivity and chemoresistance in melanoma will be tested.
The specific aims of the study are to assess the integrity of the p53 signalling pathway including downstream target genes and co-activators of p53 family members in normal melanocytes and melanoma cell lines and to relate this to cellular response to genotoxins including ultraviolet radiation (UV) and cytotoxic drugs. Experiments will first establish the existence of post-translational modifications (specifically phosphorylation and acetylation) of p53 and p73 following genotoxic stress in melanocytes and melanoma cell lines. Using p53 target gene micro-arrays the p53 transcriptosome will be analysed following genotoxic stress in primary melanocytes and melanoma cell lines of varying sensitivity to DNA damage. Expression patterns of pro-apoptotic co-activators will be monitored (in collaboration) to seek correlations between patterns of target gene expression, co-activator expression and cellular response to DNA damage.
Expression patterns of each p53 family member and their splice variants will be established in control cultures and in tissue samples obtained prospectively. The role of individual p53 family members in UV-induced apoptosis will be examined by FACS analysis by exposing p53 wild-type and p53 null melanoma cell lines to different doses of UVB, UVA and DNA-damaging agents. Experiments to upregulate (by transfection or lentiviral infection) and/or deplete p63, p73 and their splice variants (by siRNAi and micro-siRNAi) will determine the chemosensitivity of melanoma cells to relevant genotoxic stimuli. Melanoma cell lines and a spectrum of melanocytic lesions from the patient cohort will also be screened for p53 polymorphic variants to investigate their synergistic roles with p53 family members in UV-induced apoptotic cell death and correlation with clinical outcomes respectively.
The proposed work combines both functional/mechanistic studies and the molecular pathological analysis of tumour samples to assess the clinical significance of in vitro results. Determining the molecular basis of the intrinsic resistance of melanocytes to apoptosis during malignant progression may provide insight into potential therapeutic targets for overcoming their acquired chemoresistance.
The specific aims of the study are to assess the integrity of the p53 signalling pathway including downstream target genes and co-activators of p53 family members in normal melanocytes and melanoma cell lines and to relate this to cellular response to genotoxins including ultraviolet radiation (UV) and cytotoxic drugs. Experiments will first establish the existence of post-translational modifications (specifically phosphorylation and acetylation) of p53 and p73 following genotoxic stress in melanocytes and melanoma cell lines. Using p53 target gene micro-arrays the p53 transcriptosome will be analysed following genotoxic stress in primary melanocytes and melanoma cell lines of varying sensitivity to DNA damage. Expression patterns of pro-apoptotic co-activators will be monitored (in collaboration) to seek correlations between patterns of target gene expression, co-activator expression and cellular response to DNA damage.
Expression patterns of each p53 family member and their splice variants will be established in control cultures and in tissue samples obtained prospectively. The role of individual p53 family members in UV-induced apoptosis will be examined by FACS analysis by exposing p53 wild-type and p53 null melanoma cell lines to different doses of UVB, UVA and DNA-damaging agents. Experiments to upregulate (by transfection or lentiviral infection) and/or deplete p63, p73 and their splice variants (by siRNAi and micro-siRNAi) will determine the chemosensitivity of melanoma cells to relevant genotoxic stimuli. Melanoma cell lines and a spectrum of melanocytic lesions from the patient cohort will also be screened for p53 polymorphic variants to investigate their synergistic roles with p53 family members in UV-induced apoptotic cell death and correlation with clinical outcomes respectively.
The proposed work combines both functional/mechanistic studies and the molecular pathological analysis of tumour samples to assess the clinical significance of in vitro results. Determining the molecular basis of the intrinsic resistance of melanocytes to apoptosis during malignant progression may provide insight into potential therapeutic targets for overcoming their acquired chemoresistance.
Organisations
- Queen Mary, University of London, United Kingdom (Collaboration, Lead Research Organisation)
- University of Oxford, United Kingdom (Collaboration, Fellow)
- Fondazione IRCCS Istituto Nazionale dei Tumori (Collaboration)
- Imperial College London, United Kingdom (Collaboration)
- Newcastle University, United Kingdom (Collaboration)
- National Cancer Institute (CRO) (Collaboration)
- Medical Research Council (Collaboration)
- University of Dundee, United Kingdom (Collaboration)
- University of Calabria, Italy (Collaboration)
- University of Rome II (Tor Vergata), Italy (Collaboration)
- Technical University Dresden, Germany (Collaboration)
- Fondo Edo Tempia Biella (Collaboration)
- National Institute of Health and Medical Research (INSERM) (Collaboration)
- University of Cologne, Germany (Collaboration)
- University of Milan, Italy (Collaboration)
People |
ORCID iD |
Rubeta Nishat Matin (Principal Investigator / Fellow) |
Publications

Chikh A
(2011)
iASPP/p63 autoregulatory feedback loop is required for the homeostasis of stratified epithelia.
in The EMBO journal

Lambert SR
(2012)
Metastatic cutaneous squamous cell carcinoma shows frequent deletion in the protein tyrosine phosphatase receptor Type D gene.
in International journal of cancer

Lo Nigro C
(2013)
Methylated tissue factor pathway inhibitor 2 (TFPI2) DNA in serum is a biomarker of metastatic melanoma.
in The Journal of investigative dermatology

Matin RN
(2013)
Synchronous melanoma and renal carcinoma: a clinicopathological study of five cases.
in Clinical and experimental dermatology

Matin RN
(2011)
Cutaneous mucinous carcinoma arising in extramammary Paget disease of the perineum.
in The American Journal of dermatopathology

Matin RN
(2008)
Melanoma in organ transplant recipients: clinicopathological features and outcome in 100 cases.
in American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Matin RN
(2012)
KIT and BRAF mutational status in a patient with a synchronous lentigo maligna melanoma and a gastrointestinal stromal tumor.
in American journal of clinical dermatology

Matin RN
(2013)
p63 is an alternative p53 repressor in melanoma that confers chemoresistance and a poor prognosis.
in The Journal of experimental medicine

Wang H
(2012)
NT5E (CD73) is epigenetically regulated in malignant melanoma and associated with metastatic site specificity.
in British journal of cancer
Description | BSF PhD Studentship |
Amount | £81,000 (GBP) |
Funding ID | Title: Dissecting the role of iASPP, a novel crucial regulator of epidermal homeostasis, in keratinocyte skin carcinogenesis. |
Organisation | British Skin Foundation |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 09/2012 |
End | 10/2015 |
Description | Barts and the London Charitable Foundation/CRUK project grant |
Amount | £162,862 (GBP) |
Organisation | Cancer Research UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start |
Description | Barts and the London Charitable Foundation/CRUK project grant |
Amount | £99,858 (GBP) |
Organisation | Barts Charity |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start |
Description | Barts and the London Charity Research Fund |
Amount | £18,400 (GBP) |
Organisation | Barts Charity |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start |
Description | Barts and the London Charity Strategic Research Grant |
Amount | £138,800 (GBP) |
Funding ID | Title: Exploring novel molecular targets in non-BRAF mutated metastatic melanoma. |
Organisation | Barts Charity |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 01/2012 |
End | 11/2013 |
Description | Melanoma Focus Patient Impact Project |
Amount | £10,000 (GBP) |
Organisation | Melanoma Focus |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start |
Description | PhD studentship |
Amount | £75,000 (GBP) |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start |
Description | PhD studentship 2 |
Amount | £75,000 (GBP) |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start |
Description | Research Grant Award |
Amount | £15,000 (GBP) |
Organisation | The Royal Society |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start |
Title | Tissue Bank |
Description | Melanoma tissue samples and cell lines of varying stage of disease. |
Type Of Material | Database/Collection of Data/Biological Samples |
Year Produced | 2008 |
Provided To Others? | Yes |
Impact | Investigation of aberrations in other molecular pathways e.g. Wnt signalling pathway |
Description | ASPP Proteins |
Organisation | Newcastle University |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Involvement of the ASPP proteins in Autophagy |
Collaborator Contribution | Involvement of the ASPP proteins in Autophagy |
Impact | Publication listed |
Start Year | 2010 |
Description | Characterisation of melanoma |
Organisation | Fondazione IRCCS Istituto Nazionale dei Tumori |
Country | Italy |
Sector | Charity/Non Profit |
PI Contribution | We have provided the melanoma cell lines in which to determine the mutation status |
Collaborator Contribution | Established melanoma cell lines with characterized status of BRAF/NRAS mutationsGenome wide microarray analysis and microRNA profiling of melanoma cell lines silenced for p63Dissecting the role of YAP in melanomaApoptosis and DNA repair in melanoma |
Impact | Multi-disciplinary - resulted in characterisation of mutation status of melanoma lines which has led to further funding and included in publication (pending) |
Start Year | 2006 |
Description | Characterisation of melanoma |
Organisation | Fondo Edo Tempia Biella |
Country | Italy |
Sector | Charity/Non Profit |
PI Contribution | We have provided the melanoma cell lines in which to determine the mutation status |
Collaborator Contribution | Established melanoma cell lines with characterized status of BRAF/NRAS mutationsGenome wide microarray analysis and microRNA profiling of melanoma cell lines silenced for p63Dissecting the role of YAP in melanomaApoptosis and DNA repair in melanoma |
Impact | Multi-disciplinary - resulted in characterisation of mutation status of melanoma lines which has led to further funding and included in publication (pending) |
Start Year | 2006 |
Description | Characterisation of melanoma |
Organisation | National Cancer Institute (CRO) |
Country | Italy |
Sector | Academic/University |
PI Contribution | We have provided the melanoma cell lines in which to determine the mutation status |
Collaborator Contribution | Established melanoma cell lines with characterized status of BRAF/NRAS mutationsGenome wide microarray analysis and microRNA profiling of melanoma cell lines silenced for p63Dissecting the role of YAP in melanomaApoptosis and DNA repair in melanoma |
Impact | Multi-disciplinary - resulted in characterisation of mutation status of melanoma lines which has led to further funding and included in publication (pending) |
Start Year | 2006 |
Description | Characterisation of melanoma |
Organisation | University of Calabria |
Country | Italy |
Sector | Academic/University |
PI Contribution | We have provided the melanoma cell lines in which to determine the mutation status |
Collaborator Contribution | Established melanoma cell lines with characterized status of BRAF/NRAS mutationsGenome wide microarray analysis and microRNA profiling of melanoma cell lines silenced for p63Dissecting the role of YAP in melanomaApoptosis and DNA repair in melanoma |
Impact | Multi-disciplinary - resulted in characterisation of mutation status of melanoma lines which has led to further funding and included in publication (pending) |
Start Year | 2006 |
Description | Establishing a correlation between ?Np63 and IGFBP7 in melanoma cells |
Organisation | National Institute of Health and Medical Research (INSERM) |
Country | France |
Sector | Academic/University |
PI Contribution | Melanoma cell lines provision and data sharing |
Impact | Pending |
Start Year | 2010 |
Description | Serum Epigenetic Biomarkers for Metastatic Melanoma |
Organisation | University of Dundee |
Department | Centre for Oncology and Molecular Medicine |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Collaborative study investigating biomarkers in melanoma relapse - retrospective and prospective collection of samples. |
Collaborator Contribution | Collaborative study investigating biomarkers in melanoma relapse - lab research and statistical analyses undertaken at this site. |
Impact | Pending |
Start Year | 2013 |
Description | Targeting p63 |
Organisation | Imperial College London |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Cell lines, reagents and data sharing |
Collaborator Contribution | Publication listedThe role of TINT in melanoma metastasisEpigenetic regulation of novel target genes in melanomaEpigenetic regulation of novel target genes in melanoma |
Impact | Publication listed |
Start Year | 2007 |
Description | Targeting p63 |
Organisation | Queen Mary University of London |
Department | Barts and The London School of Medicine and Dentistry |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Cell lines, reagents and data sharing |
Collaborator Contribution | Publication listedThe role of TINT in melanoma metastasisEpigenetic regulation of novel target genes in melanomaEpigenetic regulation of novel target genes in melanoma |
Impact | Publication listed |
Start Year | 2007 |
Description | Targeting p63 |
Organisation | University of Cologne |
Country | Germany |
Sector | Academic/University |
PI Contribution | Cell lines, reagents and data sharing |
Collaborator Contribution | Publication listedThe role of TINT in melanoma metastasisEpigenetic regulation of novel target genes in melanomaEpigenetic regulation of novel target genes in melanoma |
Impact | Publication listed |
Start Year | 2007 |
Description | Targeting p63 |
Organisation | University of Dundee |
Department | Dundee Cancer Centre |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Cell lines, reagents and data sharing |
Collaborator Contribution | Publication listedThe role of TINT in melanoma metastasisEpigenetic regulation of novel target genes in melanomaEpigenetic regulation of novel target genes in melanoma |
Impact | Publication listed |
Start Year | 2007 |
Description | World melanoma study |
Organisation | Fondazione IRCCS Istituto Nazionale dei Tumori |
Country | Italy |
Sector | Charity/Non Profit |
PI Contribution | World melanoma study to compare multiple versus sporadic melanoma |
Impact | Pending |
Start Year | 2010 |
Description | p63 expression |
Organisation | Medical Research Council (MRC) |
Department | MRC Toxicology Unit |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Tools reagents and antibodies for detecting p63 expression in melanoma cell lines |
Collaborator Contribution | Publication pending and funding approvalTools reagents and antibodies for detecting p63 expression in melanoma cell linesCD133 expression in p63 positive melanoma cells |
Impact | Publication reported and one pending plus funding |
Start Year | 2007 |
Description | p63 expression |
Organisation | Technical University of Dresden |
Country | Germany |
Sector | Academic/University |
PI Contribution | Tools reagents and antibodies for detecting p63 expression in melanoma cell lines |
Collaborator Contribution | Publication pending and funding approvalTools reagents and antibodies for detecting p63 expression in melanoma cell linesCD133 expression in p63 positive melanoma cells |
Impact | Publication reported and one pending plus funding |
Start Year | 2007 |
Description | p63 expression |
Organisation | University of Rome Tor Vergata |
Country | Italy |
Sector | Academic/University |
PI Contribution | Tools reagents and antibodies for detecting p63 expression in melanoma cell lines |
Collaborator Contribution | Publication pending and funding approvalTools reagents and antibodies for detecting p63 expression in melanoma cell linesCD133 expression in p63 positive melanoma cells |
Impact | Publication reported and one pending plus funding |
Start Year | 2007 |
Description | p63/p53 signalling in melanoma |
Organisation | University of Dundee |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provision of melanoma cell lines, reagents and sharing of data |
Collaborator Contribution | Signalling pathwaysSignallng pathwaysp53 short isoforms and p53 Arg72Pro SNP in melanomaBak & p53 mitochondrial relocation in melanoma |
Impact | Publications listed and other outputs pending |
Start Year | 2006 |
Description | p63/p53 signalling in melanoma |
Organisation | University of Milan |
Country | Italy |
Sector | Academic/University |
PI Contribution | Provision of melanoma cell lines, reagents and sharing of data |
Collaborator Contribution | Signalling pathwaysSignallng pathwaysp53 short isoforms and p53 Arg72Pro SNP in melanomaBak & p53 mitochondrial relocation in melanoma |
Impact | Publications listed and other outputs pending |
Start Year | 2006 |
Description | p63/p53 signalling in melanoma |
Organisation | University of Oxford |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provision of melanoma cell lines, reagents and sharing of data |
Collaborator Contribution | Signalling pathwaysSignallng pathwaysp53 short isoforms and p53 Arg72Pro SNP in melanomaBak & p53 mitochondrial relocation in melanoma |
Impact | Publications listed and other outputs pending |
Start Year | 2006 |
Description | p63/p53 signalling in melanoma |
Organisation | University of Rome Tor Vergata |
Country | Italy |
Sector | Academic/University |
PI Contribution | Provision of melanoma cell lines, reagents and sharing of data |
Collaborator Contribution | Signalling pathwaysSignallng pathwaysp53 short isoforms and p53 Arg72Pro SNP in melanomaBak & p53 mitochondrial relocation in melanoma |
Impact | Publications listed and other outputs pending |
Start Year | 2006 |