teste de diagnostico molecular para câncer. metodos e composicoes sao fornecidos para identificacao de um teste de diagnostico molecular para câncer. o teste define um novo subtipo molecular deficiente de reparo de dna e permite a classificacao de um paciente dentro deste suptipo. a presente invencao pode ser usada para determinar se os pacientes com câncer sao clinicamente responsivos ou nao responsivos a um regime terapeutico antes da administracao de qualquer quimioterapia. este teste pode ser usado em tipos diferentes de câncer com diferentes drogas que afetam direta ou indiretamente dano ou reparo de dna, tal como muitas das drogas quimioterapicas citotoxicas padrao atualmente em uso. em particular, a presente invencao e dirigida ao uso de certas combinacoes de marcadores preditivos, em que a expressao dos marcadores preditivos se correlaciona com responsividade ou nao responsividade a um regime terapeutico.

Abstract BACKGROUND: We previously defined 3 molecular subgroups of High Grade Serous Ovarian Cancer (HGSOC), using gene expression data from 265 FFPE samples obtained from treatment naive patients, who received platinum based treatment following surgical resection. The 3 molecular subgroups were Angio: characterised by upregulation of angiogenesis genes; Immune: characterised by upregulation of immune genes and AngioImmune: characterised by upregulation of angiogenesis and immune genes. Patients within these 3 subgroups respond differently to standard of care treatment The Immune subgroup have the best prognosis and the Angio and AngioImmune subgroups have similar worse prognosis. A weighted gene signature to identify each of the molecular subgroups was developed. This dataset was used as a reference to investigate the effect of chemotherapy on molecular subgroup designation. METHODS: To investigate the effect of chemotherapy on predefined molecular subgroups, we analysed 35 matched pre- and post- chemotherapy samples by gene expression. The molecular subgroup assignment for each of the paired samples was determined using the gene expression signatures for each subgroup. Novel cisplatin resistant HGSOC cell lines were generated to study the mechanisms of acquired cisplatin resistance. RESULTS: 40% of the treatment naive samples that were aligned with the AngioImmune subgroup and this increased to 67.5% post-chemotherapy. 10/15 (67%) treatment naïve tumours that were initially assigned to the good prognostic Immune molecular subgroup shifted to the bad prognostic AngioImmune molecular subgroup post chemotherapy. Hence platinum chemotherapy selects for the AngioImmune subgroup, suggesting that this subgroup represents tumours which are innately platinum resistant but also provides a mechanism of acquired resistance. Additionally we demonstrate that the AngioImmune subgroup is driven by activation of the MAPK pathway and shows that cisplatin resistant HGSOC cell lines are specifically sensitive to MEK inhibitors. CONCLUSIONS: The MAPK pathway is a mechanism of innate and acquired platinum resistance in HGSOC. Furthermore the data suggests that original pre-treatment surgical/biopsy samples may fall within a different molecular subgroup to samples taken post-platinum therapy. Citation Format: Aya El-Helali, Nuala McCabe, Charlie Gourley, Andrena McCavigan, Caroline O. Michie, Bethanie Price, Niamh McGivern, Michael Churchman, Aya El-Helai, Eamonn J. O'Brien, Laura Hill, Timothy S Davison, Alistair Williams, W Glenn McCluggage, Katherine E Keating, Denis P Harkin, and Richard Kennedy. IDENTIFICATION OF A MOLECULAR SUBTYPE OF HIGH GRADE SEROUS OVARIAN CANCER REPRESENTING MAPK PATHWAY ACTIVATION AND PLATINUM RESISTANCE [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr MIP-055.

We presented univariable results according to the REMARK guidelines for associations between ColDx score and prognostic factors for recurrence-free interval (RFI; Appendix Table A1 [online only] in our article). As Casadaban et al point out, ColDx is associated with T-stage and lymphovascular invasion but not the number of nodes examined, perineural invasion, or tumor grade. It is not clear why such a relationship would be expected. The assay was designed to be independent from other known prognostic clinical factors and to add new prognostic information. As Casadaban et al suggest, we considered the subgroup of high-risk patients who we defined as exhibiting any one of the following clinical characteristics: obstruction or perforation (six patients), lymphovascular invasion (42 patients), fewer than 12 nodes sampled (176 patients), or microsatellite instability low or stable (283 patients; n 5 317; 80 RFI events). RFI was then compared between highrisk patients and low-risk patients as determined by ColDx score. Results were significant at P5 .05, with a hazard ratio of 1.62 (95%CI, 0.99 to 2.68). Thus, ColDx provides further discrimination in this higher-risk subgroup. The number of events was too small to make this comparison in the low-risk subgroup. In the parent trial, Alliance C9581, investigators sought to determine whether the use of edrecolomab—a relatively nontoxic adjuvant therapy—would demonstrate an overall survival benefit in a cohort of patients with resected, stage II colon cancer that excluded patients with high-risk factors. Patients were considered disease-free postsurgery. Thus, tumor response was not a study end point. Patient samples were obtained before treatment with edrecolomab. Overall survival and disease-free survival between treated and untreated patients were essentially equivalent (Fig 2A in our article). Nonetheless, under the case-cohort design in our validation study, we randomly selected patients stratified by assigned treatment and accounted for stratification in the analysis. Overall, toxicity was low. A maximum of grade 3 toxicity was reported for 242 (29.4%) of 823 participants who reported adverse events with edrecolomab treatment, and 48 patients (5.8%) experienced a maximum grade 4 toxicity. No individual adverse event was reported in . 5% of patients, the most prevalent of which was diarrhea. One death occurred within 30 days of completing edrecolomab therapy and was not attributed to treatment. This validation study used the same primary end point on which the gene signature was developed. Among patients who were studied in the Alliance C9581 trial, we found that it is important to distinguish between disease-related death and other causes of death in this low-risk, older patient population with stage II disease. We found large differences in outcome by sex and age for all-cause mortality that were primarily caused by association of these factors with death as a result of other causes. Including deaths as a result of other causes as an event may unduly bias results. Regarding sample insufficiency, in clinical testing, the quality control fail rate that was observed for the study is not unusual considering the average age of the formalin-fixed, paraffinembedded tissue used in the validation study (average age, 13.2 years). This limitation is acknowledged in the manuscript. In addition, average quality control fail rate within fresh formalinfixed, paraffin-embedded tissue is 5%. We stated the reason for the different prognostic score cut points in our article, which was “migration of the ColDx assay from the Affymetrix GeneChip System 3000 7G scanner to the Affymetrix microarray platform GeneChip System 3000Dx v.2.” With respect to the association with lymphocyte proliferation and activation of biologic functions with recurrence-free survival in colorectal liver metastases, the validation study was performed within primary tissuematerial. Biologic signaling within metastatic tissue is inherently different from that found within primary tissue material. That said, the most significant molecular pathways measured by the ColDx assay are detailed by Kennedy et al, among which are TGF-b and chemokine signaling, and both are associated with lymphocyte proliferation and recruitment. It is not unusual that two assays, such as the 12-gene recurrence score and ColDx, have good discrimination and calibration but do not agree with one another in individual probability predictions. It is more relevant to determine which assay is better calibrated and has better discrimination—that is, which assay is better at generating estimates that are closer to observed values. We agree with Casadaban et al that further studies are needed to demonstrate the ability of the gene expression signature to predict treatment benefit. Despite its limitations, our study was prospectively planned and used specimens and clinical data from a cohesive, well-conducted clinical trial. The results demonstrate the additive prognostic value of the measure.

Purpose Conventional staging methods are inadequate to identify patients with stage II colon cancer (CC) who are at high risk of recurrence after surgery with curative intent. ColDx is a gene expression, microarray-based assay shown to be independently prognostic for recurrence-free interval (RFI) and overall survival in CC. The objective of this study was to further validate ColDx using formalin-fixed, paraffin-embedded specimens collected as part of the Alliance phase III trial, C9581. Patients and Methods C9581 evaluated edrecolomab versus observation in patients with stage II CC and reported no survival benefit. Under an initial case-cohort sampling design, a randomly selected subcohort (RS) comprised 514 patients from 901 eligible patients with available tissue. Forty-nine additional patients with recurrence events were included in the analysis. Final analysis comprised 393 patients: 360 RS (58 events) and 33 non-RS events. Risk status was determined for each patient by ColDx. The Self-Prentice method was used to test the association between the resulting ColDx risk score and RFI adjusting for standard prognostic variables. Results Fifty-five percent of patients (216 of 393) were classified as high risk. After adjustment for prognostic variables that included mismatch repair (MMR) deficiency, ColDx high-risk patients exhibited significantly worse RFI (multivariable hazard ratio, 2.13; 95% CI, 1.3 to 3.5; P < .01). Age and MMR status were marginally significant. RFI at 5 years for patients classified as high risk was 82% (95% CI, 79% to 85%), compared with 91% (95% CI, 89% to 93%) for patients classified as low risk. Conclusion ColDx is associated with RFI in the C9581 subsample in the presence of other prognostic factors, including MMR deficiency. ColDx could be incorporated with the traditional clinical markers of risk to refine patient prognosis.

Introduction We have previously defined 3 molecular subgroups of High grade serous ovarian cancer (HGSOC), ‘Angio’, ‘Immune’ and ‘Angio_immune’ subgroups using gene expression data from 265 FFPE HGSOC samples obtained from treatment naive patients and who were subsequently treated with platinum-based standard of care (SoC) chemotherapy (carboplatin +/- paclitaxel) (Gourley, et al. J Clin Oncol 32:5s, 2014). Patients within these 3 molecular subgroups respond differently to SOC treatment. The immune subgroup has the best outcome compared to the Angio and Angio_immune subgroups (HR of 0.63 and 0.66 respectively on multivariate analysis). Since it has previously been shown that the MAPK pathway is an important mediator of cisplatin resistance in ovarian cancer, we wanted to investigate if the MAPK pathway was associated with one of the poor prognosis subgroups. Methods A gene signature that could detect each of the subgroups Angio, Immune and Angio_immune was generated from the clinical samples. Data from the Cancer Genome Atlas (TCGA) Project was used to test for correlation between each subgroup and phospho-MEK as measured by Reverse Phase Proteomic Array (RPPA). Sensitivity to the MEK inhibitor trametinib (GSK1120212) and cisplatin was determined by 10-day colony formation assay. Results We found a statistically significant association between the Angio_immune subgroup signature and Phospho-MEK (serine 217/221) expression (p = 0.047) indicating activation of the MAPK pathway in this subgroup. Additionally we have demonstrated that the Angio_immune subgroup signature is suppressed by MEK inhibition (p = 0.0055) and elevated by KRAS, NRAS and MEK1 overexpression in cell line models (0.0072, 0.0004 and Conclusion We have identified a molecular subgroup in HGSOC that is associated with MAPK signalling. A gene signature to detect this subgroup from formalin fixed paraffin embedded samples has been developed and predicts sensitivity to MEK inhibitors in pre-clinical model systems. Further work aims to validate the signature in clinical samples from patients treated with a MEK inhibitor. Citation Format: Nuala McCabe, Charlie Gourley, Andrena McGavigan, Caroline O. Michie, Niamh McGivern, Michael Churchman, Eamonn J. O’Brien, Laura Hill, Timothy S. Davison, Alistair Williams, Glenn McCluggage, Karen E. Keating, Denis P. Harkin, Richard D. Kennedy. MEK activation is associated with a molecular subgroup in high grade serous ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 453.

455 Background: Only 15-25% of pts with stage II colon cancer (CC) experience recurrence and conventional staging methods neither allow accurate identification of low (L) and high-risk (H) subgroups nor predict benefit of adjuvant chemotherapy. The ColDx assay (Almac Diagnostics) is a 634-probeset gene expression signature shown to be independently prognostic for recurrence-free interval (RFI). The objective of this study was to assess the ability of ColDx to classify stage II CC pts at L- and H-risk of relapse. Methods: This validation study was conducted using formalin fixed paraffin embedded biospecimens and clinical data from CALGB 9581, a phase III trial of edrecolomab v. observation in pts with normal risk, stage II CC. 1,454 CALGB 9,581 pts met eligibility criteria. A case-cohort sampling design was used to randomly select (RS) 514 pts from 901 eligible pts with available tissue; supplemented by 49 non-RS recurrent pts (total 563). Risk status for each pt was based on a positive or negative ColDx score using a pre-specified cutpoint, 0.4377. The Self Prentice method was used to test the association between ColDx categories and RFI (distant recurrence or death due to primary disease). Results: Initial results in 563 pts were erroneous due to a quality failure in a batch of reagent. 524 samples were re-labeled, re-ordered, and re-assayed using reagents that passed quality control (36 samples had insufficient material; 95 failed ColDx QC). Final analysis comprised 393 pts, 360 RS (58 events; 16%); 33 non-RS events. 216 pts (55%) were predicted H (62 events); 177 (45%) pts were predicted L (29 events). H pts exhibited significantly worse RFI (univariable hazard ratio (HR), 2.0; 95% CI, 1.3-3.3; p < 0.01). ColDx remained significant after adjustment for prognostic factors; HR, 2.1 (95% CI, 1.3-3.4; p < 0.01). Conclusions: The ColDx assay result is associated with RFI in the CALGB 9,581 sub-sample and is independent from other prognostic factors, including MSI. Further investigation is needed to establish the role of this classifier in guiding treatment decisions in this patient population.

BACKGROUND:There is no method routinely used to predict response to anthracycline and cyclophosphamide-based chemotherapy in the clinic; therefore patients often receive treatment for breast cancer with no benefit. Loss of the Fanconi anemia/BRCA (FA/BRCA) DNA damage response (DDR) pathway occurs in approximately 25% of breast cancer patients through several mechanisms and results in sensitization to DNA-damaging agents. The aim of this study was to develop an assay to detect DDR-deficient tumors associated with loss of the FA/BRCA pathway, for the purpose of treatment selection. METHODS:DNA microarray data from 21 FA patients and 11 control subjects were analyzed to identify genetic processes associated with a deficiency in DDR. Unsupervised hierarchical clustering was then performed using 60 BRCA1/2 mutant and 47 sporadic tumor samples, and a molecular subgroup was identified that was defined by the molecular processes represented within FA patients. A 44-gene microarray-based assay (the DDR deficiency assay) was developed to prospectively identify this subgroup from formalin-fixed, paraffin-embedded samples. All statistical tests were two-sided. RESULTS:In a publicly available independent cohort of 203 patients, the assay predicted complete pathologic response vs residual disease after neoadjuvant DNA-damaging chemotherapy (5-fluorouracil, anthracycline, and cyclophosphamide) with an odds ratio of 3.96 (95% confidence interval [Cl] =1.67 to 9.41; P = .002). In a new independent cohort of 191 breast cancer patients treated with adjuvant 5-fluorouracil, epirubicin, and cyclophosphamide, a positive assay result predicted 5-year relapse-free survival with a hazard ratio of 0.37 (95% Cl = 0.15 to 0.88; P = .03) compared with the assay negative population. CONCLUSIONS:A formalin-fixed, paraffin-embedded tissue-based assay has been developed and independently validated as a predictor of response and prognosis after anthracycline/cyclophosphamide-based chemotherapy in the neoadjuvant and adjuvant settings. These findings warrant further validation in a prospective clinical study.

PURPOSE:Current prognostic factors are poor at identifying patients at risk of disease recurrence after surgery for stage II colon cancer. Here we describe a DNA microarray-based prognostic assay using clinically relevant formalin-fixed paraffin-embedded (FFPE) samples.PATIENTS AND METHODS:A gene signature was developed from a balanced set of 73 patients with recurrent disease (high risk) and 142 patients with no recurrence (low risk) within 5 years of surgery.RESULTS:The 634-probe set signature identified high-risk patients with a hazard ratio (HR) of 2.62 (P < .001) during cross validation of the training set. In an independent validation set of 144 samples, the signature identified high-risk patients with an HR of 2.53 (P < .001) for recurrence and an HR of 2.21 (P = .0084) for cancer-related death. Additionally, the signature was shown to perform independently from known prognostic factors (P < .001).CONCLUSION:This gene signature represents a novel prognostic biomarker for patients with stage II colon cancer that can be applied to FFPE tumor samples.

OBJECTIVES:We investigated the relationship between BRCA1 protein expression by immunohistochemistry (IHC) and clinical outcome following platinum and platinum/taxane chemotherapy in sporadic epithelial ovarian cancer (EOC). METHODS:BRCA1 IHC was performed on a cohort of 292 ovarian tumours from two UK oncology centres. BRCA1 protein expression levels were correlated with overall survival (OS), progression free survival (PFS) and clinical response to chemotherapy by multivariate analysis. RESULTS:EOC patients with absent/low BRCA1 protein expression (41%) had a better chance of clinical response following chemotherapy as compared to patients with high BRCA1 expression (odds ratio 2.47: 95%CI 1.10-5.55, p=0.029). Patients with absent/low BRCA1 had a higher probability of clinical response following single agent platinum compared to high BRCA1 expressing patients (68.5% vs. 46.8%), while addition of a taxane increased response rates independent of BRCA1. Overall, patients with absent/low BRCA1 had a better clinical outcome compared to patients with high BRCA1 protein expression in terms of both OS (HR=0.65: 95%CI 0.48-0.88, p=0.006) and PFS (HR=0.74, 95%CI 0.55-0.98, p=0.040). CONCLUSIONS:We confirm that absent/low BRCA1 protein expression is a favourable prognostic marker. However, we also provide the first evidence that absent/low BRCA1 protein expression in sporadic EOC patients predicts for an improved clinical response to chemotherapy.

5000 Background: Despite histological heterogeneity and evidence of some molecular subtypes (e.g. BRCA1/2-deficient) EOC continues to be treated as a single disease. Using unsupervised hierarchical clustering of DNA microarray expression data we aimed to identify molecular subgroups with a view to personalized therapy.METHODSMicroarray expression analysis was performed on 363 FFPE EOC samples (by far the largest dataset to date) with linked prospectively collected clinical data. Unsupervised clustering was applied to the most variable genes. This was followed by computational classification in the serous samples only.RESULTSUnsupervised analysis of all samples identified 6 subgroups which were most significantly related to histology (p=3.65 x 10-27) and had different overall survival (OS; p<0.0001). Clear cell tumors were present in one group, mucinous in another, serous and endometrioid were spread across 4 groups each. When a similar approach was applied to pure serous samples (no mixed tumors; n=199) three subgroups were identified with different OS (p<0.0001). Intriguingly, the dominant discriminatory biology within the serous cohort was related to vasculature development (FDR p.vasc.dev.=7.31 x 10-7) and more specifically to angiogenic processes (FDR p.Angiogenesis= 8.59 x 10-5). Up-regulation of these angiogenesis-related genes predominated in only one of the 3 serous subgroups (58 out of 199 tumours). Class labels were assigned to these samples based upon expression of these angiogenesis-related genes and a gene expression signature was developed which could identify the novel angiogenesis molecular subgroup.CONCLUSIONS1) It is possible to consistently perform transcriptomic analysis from small FFPE ovarian tumor sections and cluster similar tumors. 2) We have identified 6 main clusters of EOC, strongly associated with histology and survival. 3) In the pure serous subgroup, 3 clusters were identified with significantly differing survival; one of these subgroups was defined by upregulation of multiple angiogenesis genes. In light of the recently reported activity of bevacizumab in EOC (GOG 218 and ICON7) this subgroup may explain the observed response.

We have developed a new method using the Qbead system for high-throughput genotyping of single nucleotide polymorphisms (SNPs). The Qbead system employs fluorescent Qdot semiconductor nanocrystals, also known as quantum dots, to encode microspheres that subsequently can be used as a platform for multiplexed assays. By combining mixtures of quantum dots with distinct emission wavelengths and intensities, unique spectral 'barcodes' are created that enable the high levels of multiplexing required for complex genetic analyses. Here, we applied the Qbead system to SNP genotyping by encoding microspheres conjugated to allele-specific oligonucleotides. After hybridization of oligonucleotides to amplicons produced by multiplexed PCR of genomic DNA, individual microspheres are analyzed by flow cytometry and each SNP is distinguished by its unique spectral barcode. Using 10 model SNPs, we validated the Qbead system as an accurate and reliable technique for multiplexed SNP genotyping. By modifying the types of probes conjugated to microspheres, the Qbead system can easily be adapted to other assay chemistries for SNP genotyping as well as to other applications such as analysis of gene expression and protein-protein interactions. With its capability for high-throughput automation, the Qbead system has the potential to be a robust and cost-effective platform for a number of applications.

Correspondence| June 21 2000 The Pro-115 → Gln mutation in peroxisome-proliferator-activated receptor (PPAR) γ2 is extremely rare in a large cohort of U.S. Caucasians Jonathan TERRETT; Jonathan TERRETT *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K. Search for other works by this author on: This Site PubMed Google Scholar John CHAMBERLAIN; John CHAMBERLAIN *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K.†Rayne Institute, Bone and Mineral Centre, Department of Medicine, Royal Free and University College Medical School, London WC1E 6JJ, U.K. Correspondence: Dr J. C. Chamberlain, Roche Products Ltd., 40 Broadwater Road, Welwyn Garden City, Herts., U.K. (e-mail chris.chamberlain@roche.com). Search for other works by this author on: This Site PubMed Google Scholar Sohaila RASTAN; Sohaila RASTAN *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K. Search for other works by this author on: This Site PubMed Google Scholar Richard MARSHALL; Richard MARSHALL *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K. Search for other works by this author on: This Site PubMed Google Scholar Ralph MCGINNIS; Ralph MCGINNIS *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K. Search for other works by this author on: This Site PubMed Google Scholar Nigel SPURR; Nigel SPURR *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K. Search for other works by this author on: This Site PubMed Google Scholar Eamonn O'BRIEN; Eamonn O'BRIEN *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K. Search for other works by this author on: This Site PubMed Google Scholar Catherine EVANS; Catherine EVANS *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K. Search for other works by this author on: This Site PubMed Google Scholar Andrew RUT Andrew RUT *Biotechnology and Genetics, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park (North), Third Avenue, Harlow, Essex CM19 5AW, U.K. Search for other works by this author on: This Site PubMed Google Scholar Clin Sci (Lond) (2000) 99 (1): 89–90. https://doi.org/10.1042/cs0990089 Article history Received: October 27 1999 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share MailTo Twitter LinkedIn Cite Icon Cite Get Permissions Citation Jonathan TERRETT, John CHAMBERLAIN, Sohaila RASTAN, Richard MARSHALL, Ralph MCGINNIS, Nigel SPURR, Eamonn O'BRIEN, Catherine EVANS, Andrew RUT; The Pro-115 → Gln mutation in peroxisome-proliferator-activated receptor (PPAR) γ2 is extremely rare in a large cohort of U.S. Caucasians. Clin Sci (Lond) 1 July 2000; 99 (1): 89–90. doi: https://doi.org/10.1042/cs0990089 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsClinical Science Search Advanced Search Keywords: obesity, pharmacogenetics, point mutation, type II diabetes mellitus, transcription factors This content is only available as a PDF. The Biochemical Society and the Medical Research Society © 20002000 Article PDF first page preview Close Modal You do not currently have access to this content.

Two forms of glycine transporter have been described to date, GlyT-1 and GlyT-2. The GlyT-2 form is expressed mainly in the spinal cord, brainstem and cerebellum. Here we describe the identification of a variant form of the human GlyT-2 (SC6), showing three amino acid changes to the previously reported protein. Population analysis identified the allele causing one of the polymorphisms, D463N, at 10% within the population with 3% being homozygous for the change. We also transfected our new variant into mammalian cells and compared it to the published cDNA, showing that the three amino acid changes present have no major effect on the biochemical properties of the transporter.