Kyle Matchett - Academia.edu (original) (raw)

Papers by Kyle Matchett

Stem cells (Dayton, Ohio), 2014

In 1953, noting a remarkable consistency between the agents causing mutations and those associate... more In 1953, noting a remarkable consistency between the agents causing mutations and those associated with cancer, Carl Nordling, a Finnish-born architect, proposed that cancer results from an accumulation of genetic mutations. It is now generally accepted that inherited mutations and environmental carcinogens can lead to the development of premalignant clones. After further mutations, one cell reaches a critical state which confers a survival or growth advantage over normal cells. Such cells have the ability to initiate a malignant tumour. They share many of the features of normal stem cells, including the capacity for self-renewal and differentiation, and are widely termed cancer stem cells (CSCs). Although CSCs have been well characterized in hematological malignancies, their existence in some other tissues has been questioned. Here, we review recent work in which stem cells and stem cell-like cells have been used to investigate the pathogenesis of cancer and potential anticancer tr...

British journal of haematology, 2015

Recombinant human erythropoietin (rHuEPO) is an effective treatment for anaemia but concerns that... more Recombinant human erythropoietin (rHuEPO) is an effective treatment for anaemia but concerns that it causes disease progression in cancer patients by activation of EPO receptors (EPOR) in tumour tissue have been controversial and have restricted its clinical use. Initial clinical studies were flawed because they used polyclonal antibodies, later shown to lack specificity for EPOR. Moreover, multiple isoforms of EPOR caused by differential splicing have been reported in cancer cell lines at the mRNA level but investigations of these variants and their potential impact on tumour progression, have been hampered by lack of suitable antibodies. The EpoCan consortium seeks to promote improved pathological testing of EPOR, leading to safer clinical use of rHuEPO, by producing well characterized EPOR antibodies. Using novel genetic and traditional peptide immunization protocols, we have produced mouse and rat monoclonal antibodies, and show that several of these specifically recognize EPOR ...

Advances in experimental medicine and biology, 2014

Ran is a small ras-related GTPase that controls the nucleocytoplasmic exchange of macromolecules ... more Ran is a small ras-related GTPase that controls the nucleocytoplasmic exchange of macromolecules across the nuclear envelope. It binds to chromatin early during nuclear formation and has important roles during the eukaryotic cell cycle, where it regulates mitotic spindle assembly, nuclear envelope formation and cell cycle checkpoint control. Like other GTPases, Ran relies on the cycling between GTP-bound and GDP-bound conformations to interact with effector proteins and regulate these processes. In nucleocytoplasmic transport, Ran shuttles across the nuclear envelope through nuclear pores. It is concentrated in the nucleus by an active import mechanism where it generates a high concentration of RanGTP by nucleotide exchange. It controls the assembly and disassembly of a range of complexes that are formed between Ran-binding proteins and cellular cargo to maintain rapid nuclear transport. Ran also has been identified as an essential protein in nuclear envelope formation in eukaryotes...

Cancer Research

A common feature of the septin family of genes is alternative splicing with generation of multipl... more A common feature of the septin family of genes is alternative splicing with generation of multiple transcripts. In SEPT9, up to 18 such transcripts have been identified by various combinations of 5’ and 3’ ends. With respect to the 5’ transcripts, we have previously reported that two of these, SEPT9_v1 and v4* are upregulated in sporadic ovarian tumors. Our current studies are focused on determining the significance of this observation. Initially, we sought to determine if these changes are clinically relevant. We therefore analyzed SEPT9 expression in a bank of ovarian tumors for which outcome and various clinical parameters were known. RNA was extracted from 100 FFPE ovarian tumor sections from women enrolled in the Scottish Randomised Trial in Ovarian Cancer (SCOTROC), which compares paclitaxel-carboplatin and docetaxel-carboplatin chemotherapy regimes. Primers were optimized to amplify each 5’ SEPT9 transcript by qRT-PCR in each sample and the profile analyzed for correlations w...

Objectives: Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ova... more Objectives: Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ovarian cancers. The BRCA1 protein functions to maintain genomic stability via important roles in DNA repair, transcriptional regulation, and post-replicative repair. Despite functions in processes essential in all cells, BRCA1 loss or mutation leads to tumours predominantly in estrogen-regulated tissues. Here, we aim to determine if endogenous estrogen metabolites may be an initiator of genomic instability in BRCA1 deficient cells. Methods: We analysed DNA DSBs by γH2AX, 53BP1, and pATM1981 foci and neutral comet assay, estrogen metabolite concentrations by LC-MS/MS, and BRCA1 transcriptional regulation of metabolism genes by ChIP-chip, ChIP, and qRT-PCR. Results: We show that estrogen metabolism is perturbed in BRCA1 deficient cells resulting in elevated production of 2-hydroxyestradiol (2-OHE2) and 4-hydroxyestradiol (4-OHE2), and decreased production of the protective metabolite 4-metho...

Current Oncology, 2012

Background: Nearly 15% of DNA tests for BRCA1/2 results in the identification of an unclassified ... more Background: Nearly 15% of DNA tests for BRCA1/2 results in the identification of an unclassified variant (UV). In DNA diagnostic laboratories in The Netherlands, a 4-group classification system (class I to IV ) is in use (Bell et al.). Aim of this study was to investigate whether the UVs in different classes showed a significant difference in their in silico characteristics and would justify current differences in protocols for counselling with respect to communication to the counselees.

Expert Review of Molecular Diagnostics, 2013

A vast body of research in breast cancer prognostication has accumulated. Yet despite this, patie... more A vast body of research in breast cancer prognostication has accumulated. Yet despite this, patients within current prognostic categories may have significantly different outcomes. There is a need to more accurately divide those cancer types associated with an excellent prognosis from those requiring more aggressive therapy. Gene expression array studies have revealed the numerous molecular breast cancer subtypes that are associated with differing outcomes. Furthermore, as next generation technologies evolve and further reveal the complexities of breast cancer, it is likely that existing prognostic approaches will become progressively refined. Future prognostication in breast cancer requires a morphomolecular, multifaceted approach involving the assessment of anatomical disease extent and levels of protein, DNA and RNA expression. One of the major challenges in prognostication will be the integration of potential assays into existing clinical systems and identification of appropriate patient subgroups for analysis.

Cancer Research, 2014

Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ovarian cancers... more Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ovarian cancers. BRCA1 functions to maintain genomic stability through critical roles in DNA repair, cell-cycle arrest, and transcriptional control. A major question has been why BRCA1 loss or mutation leads to tumors mainly in estrogen-regulated tissues, given that BRCA1 has essential functions in all cell types. Here, we report that estrogen and estrogen metabolites can cause DNA double-strand breaks (DSB) in estrogen receptora-negative breast cells and that BRCA1 is required to repair these DSBs to prevent metabolite-induced genomic instability. We found that BRCA1 also regulates estrogen metabolism and metabolite-mediated DNA damage by repressing the transcription of estrogen-metabolizing enzymes, such as CYP1A1, in breast cells. Finally, we used a knock-in human cell model with a heterozygous BRCA1 pathogenic mutation to show how BRCA1 haploinsufficiency affects these processes. Our findings provide pivotal new insights into why BRCA1 mutation drives the formation of tumors in estrogen-regulated tissues, despite the general role of BRCA1 in DNA repair in all cell types. Cancer Res; 74(10); 2773-84. Ó2014 AACR.

Cancer Research, 2011

5012: Regulation of splicing of SEPT9 in health and disease Abstract 5012: Regulation of splicing... more 5012: Regulation of splicing of SEPT9 in health and disease Abstract 5012: Regulation of splicing of SEPT9 in health and disease

Cancer Research, 2010

260: Elucidating the roles of the alternatively spliced transcripts of Septin 9

Cytokine & Growth Factor Reviews, 2015

Breast cancer is one of the most prevalent malignancies worldwide. It consists of a group of tumo... more Breast cancer is one of the most prevalent malignancies worldwide. It consists of a group of tumor cells that have the ability to grow uncontrollably, overcome replicative senescence (tumor progression) and metastasize within the body. Metastases are processes that consist of an array of complex gene dysregulation events. Although these processes are still not fully understood, the dysregulation of a number of key proteins must take place if the tumor cells are to disseminate and metastasize. It is now widely accepted that future effective and innovative treatments of cancer metastasis will have to encompass all the major components of malignant transformation. For this reason, much research is now being carried out into the mechanisms that govern the malignant transformation processes. Recent research has identified key genes involved in the development of metastases, as well as their mechanisms of action. A detailed understanding of the encoded proteins and their interrelationship generates the possibility of developing novel therapeutic approaches. This review will focus on a select group of proteins, often deregulated in breast cancer metastasis, which have shown therapeutic promise, notably, EMT, E-cadherin, Osteopontin, PEA3, Transforming Growth Factor Beta (TGF-b) and Ran.

Stem cells (Dayton, Ohio), 2014

In 1953, noting a remarkable consistency between the agents causing mutations and those associate... more In 1953, noting a remarkable consistency between the agents causing mutations and those associated with cancer, Carl Nordling, a Finnish-born architect, proposed that cancer results from an accumulation of genetic mutations. It is now generally accepted that inherited mutations and environmental carcinogens can lead to the development of premalignant clones. After further mutations, one cell reaches a critical state which confers a survival or growth advantage over normal cells. Such cells have the ability to initiate a malignant tumour. They share many of the features of normal stem cells, including the capacity for self-renewal and differentiation, and are widely termed cancer stem cells (CSCs). Although CSCs have been well characterized in hematological malignancies, their existence in some other tissues has been questioned. Here, we review recent work in which stem cells and stem cell-like cells have been used to investigate the pathogenesis of cancer and potential anticancer tr...

British journal of haematology, 2015

Recombinant human erythropoietin (rHuEPO) is an effective treatment for anaemia but concerns that... more Recombinant human erythropoietin (rHuEPO) is an effective treatment for anaemia but concerns that it causes disease progression in cancer patients by activation of EPO receptors (EPOR) in tumour tissue have been controversial and have restricted its clinical use. Initial clinical studies were flawed because they used polyclonal antibodies, later shown to lack specificity for EPOR. Moreover, multiple isoforms of EPOR caused by differential splicing have been reported in cancer cell lines at the mRNA level but investigations of these variants and their potential impact on tumour progression, have been hampered by lack of suitable antibodies. The EpoCan consortium seeks to promote improved pathological testing of EPOR, leading to safer clinical use of rHuEPO, by producing well characterized EPOR antibodies. Using novel genetic and traditional peptide immunization protocols, we have produced mouse and rat monoclonal antibodies, and show that several of these specifically recognize EPOR ...

Advances in experimental medicine and biology, 2014

Ran is a small ras-related GTPase that controls the nucleocytoplasmic exchange of macromolecules ... more Ran is a small ras-related GTPase that controls the nucleocytoplasmic exchange of macromolecules across the nuclear envelope. It binds to chromatin early during nuclear formation and has important roles during the eukaryotic cell cycle, where it regulates mitotic spindle assembly, nuclear envelope formation and cell cycle checkpoint control. Like other GTPases, Ran relies on the cycling between GTP-bound and GDP-bound conformations to interact with effector proteins and regulate these processes. In nucleocytoplasmic transport, Ran shuttles across the nuclear envelope through nuclear pores. It is concentrated in the nucleus by an active import mechanism where it generates a high concentration of RanGTP by nucleotide exchange. It controls the assembly and disassembly of a range of complexes that are formed between Ran-binding proteins and cellular cargo to maintain rapid nuclear transport. Ran also has been identified as an essential protein in nuclear envelope formation in eukaryotes...

Cancer Research

A common feature of the septin family of genes is alternative splicing with generation of multipl... more A common feature of the septin family of genes is alternative splicing with generation of multiple transcripts. In SEPT9, up to 18 such transcripts have been identified by various combinations of 5’ and 3’ ends. With respect to the 5’ transcripts, we have previously reported that two of these, SEPT9_v1 and v4* are upregulated in sporadic ovarian tumors. Our current studies are focused on determining the significance of this observation. Initially, we sought to determine if these changes are clinically relevant. We therefore analyzed SEPT9 expression in a bank of ovarian tumors for which outcome and various clinical parameters were known. RNA was extracted from 100 FFPE ovarian tumor sections from women enrolled in the Scottish Randomised Trial in Ovarian Cancer (SCOTROC), which compares paclitaxel-carboplatin and docetaxel-carboplatin chemotherapy regimes. Primers were optimized to amplify each 5’ SEPT9 transcript by qRT-PCR in each sample and the profile analyzed for correlations w...

Objectives: Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ova... more Objectives: Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ovarian cancers. The BRCA1 protein functions to maintain genomic stability via important roles in DNA repair, transcriptional regulation, and post-replicative repair. Despite functions in processes essential in all cells, BRCA1 loss or mutation leads to tumours predominantly in estrogen-regulated tissues. Here, we aim to determine if endogenous estrogen metabolites may be an initiator of genomic instability in BRCA1 deficient cells. Methods: We analysed DNA DSBs by γH2AX, 53BP1, and pATM1981 foci and neutral comet assay, estrogen metabolite concentrations by LC-MS/MS, and BRCA1 transcriptional regulation of metabolism genes by ChIP-chip, ChIP, and qRT-PCR. Results: We show that estrogen metabolism is perturbed in BRCA1 deficient cells resulting in elevated production of 2-hydroxyestradiol (2-OHE2) and 4-hydroxyestradiol (4-OHE2), and decreased production of the protective metabolite 4-metho...

Current Oncology, 2012

Background: Nearly 15% of DNA tests for BRCA1/2 results in the identification of an unclassified ... more Background: Nearly 15% of DNA tests for BRCA1/2 results in the identification of an unclassified variant (UV). In DNA diagnostic laboratories in The Netherlands, a 4-group classification system (class I to IV ) is in use (Bell et al.). Aim of this study was to investigate whether the UVs in different classes showed a significant difference in their in silico characteristics and would justify current differences in protocols for counselling with respect to communication to the counselees.

Expert Review of Molecular Diagnostics, 2013

A vast body of research in breast cancer prognostication has accumulated. Yet despite this, patie... more A vast body of research in breast cancer prognostication has accumulated. Yet despite this, patients within current prognostic categories may have significantly different outcomes. There is a need to more accurately divide those cancer types associated with an excellent prognosis from those requiring more aggressive therapy. Gene expression array studies have revealed the numerous molecular breast cancer subtypes that are associated with differing outcomes. Furthermore, as next generation technologies evolve and further reveal the complexities of breast cancer, it is likely that existing prognostic approaches will become progressively refined. Future prognostication in breast cancer requires a morphomolecular, multifaceted approach involving the assessment of anatomical disease extent and levels of protein, DNA and RNA expression. One of the major challenges in prognostication will be the integration of potential assays into existing clinical systems and identification of appropriate patient subgroups for analysis.

Cancer Research, 2014

Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ovarian cancers... more Germline mutations in BRCA1 predispose carriers to a high incidence of breast and ovarian cancers. BRCA1 functions to maintain genomic stability through critical roles in DNA repair, cell-cycle arrest, and transcriptional control. A major question has been why BRCA1 loss or mutation leads to tumors mainly in estrogen-regulated tissues, given that BRCA1 has essential functions in all cell types. Here, we report that estrogen and estrogen metabolites can cause DNA double-strand breaks (DSB) in estrogen receptora-negative breast cells and that BRCA1 is required to repair these DSBs to prevent metabolite-induced genomic instability. We found that BRCA1 also regulates estrogen metabolism and metabolite-mediated DNA damage by repressing the transcription of estrogen-metabolizing enzymes, such as CYP1A1, in breast cells. Finally, we used a knock-in human cell model with a heterozygous BRCA1 pathogenic mutation to show how BRCA1 haploinsufficiency affects these processes. Our findings provide pivotal new insights into why BRCA1 mutation drives the formation of tumors in estrogen-regulated tissues, despite the general role of BRCA1 in DNA repair in all cell types. Cancer Res; 74(10); 2773-84. Ó2014 AACR.

Cancer Research, 2011

5012: Regulation of splicing of SEPT9 in health and disease Abstract 5012: Regulation of splicing... more 5012: Regulation of splicing of SEPT9 in health and disease Abstract 5012: Regulation of splicing of SEPT9 in health and disease

Cancer Research, 2010

260: Elucidating the roles of the alternatively spliced transcripts of Septin 9

Cytokine & Growth Factor Reviews, 2015

Breast cancer is one of the most prevalent malignancies worldwide. It consists of a group of tumo... more Breast cancer is one of the most prevalent malignancies worldwide. It consists of a group of tumor cells that have the ability to grow uncontrollably, overcome replicative senescence (tumor progression) and metastasize within the body. Metastases are processes that consist of an array of complex gene dysregulation events. Although these processes are still not fully understood, the dysregulation of a number of key proteins must take place if the tumor cells are to disseminate and metastasize. It is now widely accepted that future effective and innovative treatments of cancer metastasis will have to encompass all the major components of malignant transformation. For this reason, much research is now being carried out into the mechanisms that govern the malignant transformation processes. Recent research has identified key genes involved in the development of metastases, as well as their mechanisms of action. A detailed understanding of the encoded proteins and their interrelationship generates the possibility of developing novel therapeutic approaches. This review will focus on a select group of proteins, often deregulated in breast cancer metastasis, which have shown therapeutic promise, notably, EMT, E-cadherin, Osteopontin, PEA3, Transforming Growth Factor Beta (TGF-b) and Ran.