- Email: [email protected]
MC13-0065 Quantitative assessment of VEGF, BCL6 and CMYC predicts outcome in patients with DLBCL treated with R-CHOP: Implications for guiding future treatment selection
S30 a histological review of 510 cases. In addition, 13 cases of a subset of follicular adenomatoid nodules with focal areas showing nuclear features characteristic of PTC, identified as putative PTC precursor lesion, were also analyzed. Immunohistochemical analysis of galectin-3, HBME-1, CK 19 and the proliferation markers Ki 67 and cyclin D1 was performed. Lesions were analyzed for cyclin D1 gene amplification by fluorescent in situ hybridization. PTC also frequently carries several genetic alterations in genes coding for proteins that activate the mitogen-activated protein kinases (MAPK) signaling pathway, which plays a key role in the regulation of cell growth and differentiation. The role of MAPK pathway activity in PTC was investigated by immunohistochemical labelling of phosphorylated ERK, JNK and p38. Results: All putative precursor lesions showed immunolabelling of cyclin D1 and Ki 67; 11/13 cases showed immunolabelling of CK 19; 10/13 cases showed immunolabelling of HBME-1 and 4/13 cases showed immunolabelling of galectin-3. Surrounding adenomatoid areas showed no to faint focal staining of cyclin D1, HBME-1 and galectin-3 in all thirteen cases. A low rate of cyclin D1 gene amplification (Figure) was identified in a significant proportion of cells in the putative precursor lesion as compared to surrounding benign adenomatoid areas. ERK and JNK activation was seen in 50 and 35 percent of PTC cases with immunolabelling in less than 10 percent of cells. p38 MAPK phosphorylation was seen as abundant cytoplasmic immunolabelling in 55% of PTC cases and 60% of putative precursor lesion cases. A one way ANOVA test showed significant difference between the ERK, JNK and p38 phosphorylation (p<0.01).
Poster Presentations (GEP) to sub-classify patients and predict outcome. Although promising, these assays require fresh frozen tissue and for most health centers remain prohibitively out of reach due to cost. Purpose/Objective: We sought to confirm our original observation that increasing levels of VEGF and BCL6 were associated with reduced outcome using an expanded patient cohort evaluated with quantitative immunofluorescence (QIF). In addition, we wanted to assess whether QIF, using biomarkers currently employed in the Colomo and Hans algorithm, is comparable to GEP data, for risk stratification. Materials and Methods: Complete clinical and QIF data for 118 patients from three different sites in Spain were evaluated to predict response to standard R-CHOP. Multiplex QIF with CRI Nuance imaging software was performed for selected markers including: MUM1, CD10, CD20, BCL6, BCL2, VEGF, and CMYC. The Colomo and Hans algorithm’s using QIF were compared with previously obtained GEP data on 36 patients classified as either ABC (n=14) or GCB (n=22). Kaplan-Meier survival function curves, concordance index (CoI) and multivariate models were employed to associate marker expression with OS and PFS. Results: For all three cohorts, both low IPI and female gender predicted good overall survival (p<0.001, respectively). We validated our previous VEGF cutpoint and confirmed high levels were associated with poor PFS (0.042). We then re-calibrated the VEGF cut-point on 118 patients and further improved this association for both OS (p=0.003) and PFS (p=0.004). We also validated our prior BCL6 cut-point and confirmed high levels were associated with poor PFS (p=0.001) and after re-calibration the results were even more significant (OS, p=0.008; PFS, p=0.001). In addition, increasing amounts of CMYC were associated with poor PFS and OS, respectively (p=0.014, 0.021). Mum1 by QIF alone was able to risk stratify patients classified by GEP (p<0.05) suggesting a role for IF and other markers in future algorithms. Best multivariate model to predict OS or PFS included the IPI score and BCL6. Clinical heterogeneity impacted on selected marker performance between cohorts. Conclusions: VEGF and BCL6 may be useful markers for predicting likely patient-specific response to current therapies. QIF with additional biomarkers and integrated with clinical data should be comparable with GEP to risk stratify patients in the future.
MC13-0066 ORILAB a functional and molecular imaging corelab for cancer research
Figure 1. Amplification of cyclin D1 gene seen in the putative precursor lesion using FISH. Cyclin D1 gene appears in red whereas chromosome centromeric repeat region is green (bar = 20 μm).
Conclusions: Increased expression of cyclin D1 and amplification of its gene along with immunolabelling of HBME-1 and p38 phosphorylation in areas showing cytological features of PTC within follicular adenomatoid nodules suggest that these areas could correspond to a precursor lesion of follicular variant of PTC. Increased expression of p38-MAPK cascade in PTC variants indicate that it is functional in PTC. p38-MAPK hyper-expression in the precursor lesion can act as a potential complementary marker. However, its role in the tumorigenesis of PTC needs to elucidated.
MC13-0065 Quantitative assessment of VEGF, BCL6 and CMYC predicts outcome in patients with DLBCL treated with R-CHOP: Implications for guiding future treatment selection M. Donovan 1 , N. Erill 2 , P. Puig 2 , A. Colomer 2 , L. Colomo 3 , E. Campo 4 . 1 Pathology, Mt. Sinai Medical Center, New York, USA; 2 Pathology, Althia Health S.L., Barcelona, Spain; 3 Pathology, Hospital Clinic, Barcelona, Spain; 4 Pathology, Hospital Clinic, University of Barcelona, Barcelona, Spain Background: The pathogenesis of DLBCL is both complex and heterogeneous, and pathogenetic mechanisms remain largely unknown. Although the addition of Rituximab (R) to CHOP has significantly improved outcome, approximately 30–40% of patients ultimately die of their disease supporting the need to improve risk stratification and understand response to therapy. The international prognostic index (IPI) remains the “gold standard” for progression free survival (PFS) and overall survival (OS); however, more recently, several groups have reported the utility of gene expression profiles
L. Belenguer-Querol 1 , T. Guiot 2 , C. Garcia 2 , S. Chao 3 , S. Drisis 3 , M. Lemort 3 , P. Flamen 2 . 1 Oncology Related Imaging coreLAB, Institut Jules Bordet, Brussels, Belgium; 2 Nuclear Medicine, Institut Jules Bordet, Brussels, Belgium; 3 Radiology, Institut Jules Bordet, Brussels, Belgium Background: Functional and molecular imaging techniques allow the identification of additional cancer imaging biomarkers for prognosis or prediction of patient response to a specific treatment. Biomarkers technical feasibility and diagnostic accuracy are available yet, however large scale multicentric trials are required to validate them before implementation in clinical routine. Setting up these studies involving functional and molecular imaging becomes cumbersome due to image acquisition intra and inter-center variability and differences in image quality and analysis. Purpose/Objective: An Oncology Related Imaging coreLAB platform, ORILAB has been established within a university dedicated cancer hospital environment to facilitate the use of imaging biomarkers in multicentric studies through standardization and harmonization of imaging data acquisition, analysis and storage. Materials and Methods: ORILAB exploits available in-house academic resources from multiple disciplines including physicists, engineers and physicians. The platform entails definition of imaging protocols, a Clinical Data Management System (CDMS) to electronically capture imaging trial data from participating sites or from image reviewers, a Quality Management System (QMS) to assure quality of source data and a centralized review (CR) for image interpretation. The CR is constituted by Nuclear Medicine and Radiology physicians with extensive experience in the development of new cancer biomarkers and with a special interest in translating these imaging procedures into clinical trials which demands greater standardization endeavors. Results: Imaging protocols describe guidelines for standardized acquisition and transmission of imaging data reducing intra and inter-center variability. Electronic encoding of imaging trial data results in faster and more reliable input of trial data in multicentric studies by an earlier detection of noncompliance and queries. The QMS allows the adaptation of the CDMS workflow to
Poster Presentations (GEP) to sub-classify patients and predict outcome. Although promising, these assays require fresh frozen tissue and for most health centers remain prohibitively out of reach due to cost. Purpose/Objective: We sought to confirm our original observation that increasing levels of VEGF and BCL6 were associated with reduced outcome using an expanded patient cohort evaluated with quantitative immunofluorescence (QIF). In addition, we wanted to assess whether QIF, using biomarkers currently employed in the Colomo and Hans algorithm, is comparable to GEP data, for risk stratification. Materials and Methods: Complete clinical and QIF data for 118 patients from three different sites in Spain were evaluated to predict response to standard R-CHOP. Multiplex QIF with CRI Nuance imaging software was performed for selected markers including: MUM1, CD10, CD20, BCL6, BCL2, VEGF, and CMYC. The Colomo and Hans algorithm’s using QIF were compared with previously obtained GEP data on 36 patients classified as either ABC (n=14) or GCB (n=22). Kaplan-Meier survival function curves, concordance index (CoI) and multivariate models were employed to associate marker expression with OS and PFS. Results: For all three cohorts, both low IPI and female gender predicted good overall survival (p<0.001, respectively). We validated our previous VEGF cutpoint and confirmed high levels were associated with poor PFS (0.042). We then re-calibrated the VEGF cut-point on 118 patients and further improved this association for both OS (p=0.003) and PFS (p=0.004). We also validated our prior BCL6 cut-point and confirmed high levels were associated with poor PFS (p=0.001) and after re-calibration the results were even more significant (OS, p=0.008; PFS, p=0.001). In addition, increasing amounts of CMYC were associated with poor PFS and OS, respectively (p=0.014, 0.021). Mum1 by QIF alone was able to risk stratify patients classified by GEP (p<0.05) suggesting a role for IF and other markers in future algorithms. Best multivariate model to predict OS or PFS included the IPI score and BCL6. Clinical heterogeneity impacted on selected marker performance between cohorts. Conclusions: VEGF and BCL6 may be useful markers for predicting likely patient-specific response to current therapies. QIF with additional biomarkers and integrated with clinical data should be comparable with GEP to risk stratify patients in the future.
MC13-0066 ORILAB a functional and molecular imaging corelab for cancer research
Figure 1. Amplification of cyclin D1 gene seen in the putative precursor lesion using FISH. Cyclin D1 gene appears in red whereas chromosome centromeric repeat region is green (bar = 20 μm).
Conclusions: Increased expression of cyclin D1 and amplification of its gene along with immunolabelling of HBME-1 and p38 phosphorylation in areas showing cytological features of PTC within follicular adenomatoid nodules suggest that these areas could correspond to a precursor lesion of follicular variant of PTC. Increased expression of p38-MAPK cascade in PTC variants indicate that it is functional in PTC. p38-MAPK hyper-expression in the precursor lesion can act as a potential complementary marker. However, its role in the tumorigenesis of PTC needs to elucidated.
MC13-0065 Quantitative assessment of VEGF, BCL6 and CMYC predicts outcome in patients with DLBCL treated with R-CHOP: Implications for guiding future treatment selection M. Donovan 1 , N. Erill 2 , P. Puig 2 , A. Colomer 2 , L. Colomo 3 , E. Campo 4 . 1 Pathology, Mt. Sinai Medical Center, New York, USA; 2 Pathology, Althia Health S.L., Barcelona, Spain; 3 Pathology, Hospital Clinic, Barcelona, Spain; 4 Pathology, Hospital Clinic, University of Barcelona, Barcelona, Spain Background: The pathogenesis of DLBCL is both complex and heterogeneous, and pathogenetic mechanisms remain largely unknown. Although the addition of Rituximab (R) to CHOP has significantly improved outcome, approximately 30–40% of patients ultimately die of their disease supporting the need to improve risk stratification and understand response to therapy. The international prognostic index (IPI) remains the “gold standard” for progression free survival (PFS) and overall survival (OS); however, more recently, several groups have reported the utility of gene expression profiles
L. Belenguer-Querol 1 , T. Guiot 2 , C. Garcia 2 , S. Chao 3 , S. Drisis 3 , M. Lemort 3 , P. Flamen 2 . 1 Oncology Related Imaging coreLAB, Institut Jules Bordet, Brussels, Belgium; 2 Nuclear Medicine, Institut Jules Bordet, Brussels, Belgium; 3 Radiology, Institut Jules Bordet, Brussels, Belgium Background: Functional and molecular imaging techniques allow the identification of additional cancer imaging biomarkers for prognosis or prediction of patient response to a specific treatment. Biomarkers technical feasibility and diagnostic accuracy are available yet, however large scale multicentric trials are required to validate them before implementation in clinical routine. Setting up these studies involving functional and molecular imaging becomes cumbersome due to image acquisition intra and inter-center variability and differences in image quality and analysis. Purpose/Objective: An Oncology Related Imaging coreLAB platform, ORILAB has been established within a university dedicated cancer hospital environment to facilitate the use of imaging biomarkers in multicentric studies through standardization and harmonization of imaging data acquisition, analysis and storage. Materials and Methods: ORILAB exploits available in-house academic resources from multiple disciplines including physicists, engineers and physicians. The platform entails definition of imaging protocols, a Clinical Data Management System (CDMS) to electronically capture imaging trial data from participating sites or from image reviewers, a Quality Management System (QMS) to assure quality of source data and a centralized review (CR) for image interpretation. The CR is constituted by Nuclear Medicine and Radiology physicians with extensive experience in the development of new cancer biomarkers and with a special interest in translating these imaging procedures into clinical trials which demands greater standardization endeavors. Results: Imaging protocols describe guidelines for standardized acquisition and transmission of imaging data reducing intra and inter-center variability. Electronic encoding of imaging trial data results in faster and more reliable input of trial data in multicentric studies by an earlier detection of noncompliance and queries. The QMS allows the adaptation of the CDMS workflow to