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Identification of candidate predictive protein biomarkers by M2 proteomics for clinical onset and treatment efficacy of multiple sclerosis
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Abstracts
Objective: Autoantibodies targeting aquaporin-4 (AQP4) are an important biomarker in neuromyelitis optica. Furthermore, autoantibodies to myelin oligodendrocyte glycoprotein (MOG) are detected in pediatric demyelinating disorders. We examined a cohort of adults with AQP4 antibody-negative neuromyelitis optica spectrum disorder (NMOSD) for antibodies to MOG. Methods: We performed a flow cytometry cell-based assay using live human embryonic kidney 293 cells that were lentivirus-transduced cells to express full-length surface MOG. Serum was tested in 23 AQP4 antibody-negative NMOSD patients with bilateral and/or recurrent optic neuritis (BON, n = 11), longitudinally extensive transverse myelitis (LETM, n = 10), and sequential BON and LETM (n = 2). Control cohorts included patients with clinically definite multiple sclerosis (MS, n = 76), as well as age matched healthy and other neurological disease controls (n = 52). Results: MOG antibodies were detected in 9/23 AQP4 antibodynegative NMOSD patients compared to 1/76 MS patients and 0/52 controls (P b 0.001). In all patients, the MOG antibodies were of the IgG rather than the IgM isotype. MOG antibodies were detected in 8/11 BON, 0/10 LETM, and 1/2 sequential BON and LETM patients. 6/9 MOG antibody-positive AQP4 antibody-negative patients had a relapsing course. MOG antibody-positive patients were more often female, of younger age at disease onset, and had a preceding viral prodrome compared to MOG antibody-negative patients (P values not significant). MOG antibody-positive patients had prominent bilateral optic disc swelling at presentation, and were more likely to have a rapid response to steroid therapy and relapse on steroid cessation than MOG antibody-negative patients (P = 0.034, P = 0.029, respectively). While 8/9 MOG antibody-positive patients had good follow-up visual acuity, one experienced sustained impairments in visual acuity and visual field testing. Furthermore, three patients had retinal nerve fiber layer atrophy on optical coherence tomography at follow-up, and one had residual spinal disability. Conclusions: MOG antibodies have a strong association with BON and may be a useful clinical biomarker. MOG antibody-associated BON is a relapsing disorder that is frequently steroid responsive and often steroid dependent. Failure of early recognition and institution of immunotherapy may be associated with sustained impairment. doi:10.1016/j.jneuroim.2014.08.065
113 Identification of candidate predictive protein biomarkers by M2 proteomics for clinical onset and treatment efficacy of multiple sclerosis Itay Raphael, Thomas G. Forsthuber The University of Texas at San Antonio, The University of Texas at San Antonio, San Antonio, United States Despite extensive research, multiple sclerosis (MS) remains a disease that lacks a definitive diagnostic test to predict imminent disease relapses. Thus, patients may undergo years of unnecessary treatments. Additionally, current treatments for MS can produce dramatically different outcomes in different individuals and therefore there is a critical need to develop biomarkers for treatment efficacy and resistance. We have recently developed a novel quantitative Microwave & Magnetic (M2) proteomics method to quantitatively measure changes in proteome expression over the course of experimental autoimmune encephalomyelitis (EAE), the standard murine animal model of MS. Our statistical analyses, including protein trajectories and receiver operating characteristic
(ROC) curves, indicate a strong correlation to EAE severity, and/or clinical-phase (i.e. time). Interestingly, M2 proteomics revealed characteristic CNS-specific protein expression waves prior to the onset of clinical symptoms. We are currently testing whether these characteristic protein expression waves allow us to predict the onset of clinical symptoms and forecast the severity of the disease. Furthermore, we have identified changes in the CNS proteome during EAE that correlate with the therapeutic efficacy of glucocorticoid treatment. Our studies will provide proof-of-principle for developing homologous human biomarkers that may be useful to predict disease onset, clinical severity and treatment efficacy. Finally, the changes in the CNS proteome detected by M2 proteomics may provide insights into key mechanisms that contribute to the disease pathology and may be useful to develop new therapeutic targets for MS. doi:10.1016/j.jneuroim.2014.08.066
495 JCV index and L-selectin for natalizumab-associated PML risk stratification Nicholas Schwab, Tilman Schneider-Hohendorf, Johanna Breuer, Anita Posevitz-Fejfar, Heinz Wiendl Neurology, University of Muenster, Muenster, Germany Background: Long-term treatment with and the presence of anti-JCV antibodies in serum are associated with the risk to develop-induced PML. Recent data suggest that the level of anti-JCV antibodies in serum (JCV antibody index N 0.9) and/or the lack of L-selectin (CD62L) on cryopreserved CD4+ T cells (%CD62L + cells of CD4+ T cells b 21.6) could be biomarkers for higher PML risk. Objectives: To compare and correlate PML risk stratification parameters during natalizumab therapy including the assessment of their relationship. Methods: Up to 1921 patients (number depending on data set completion) were analyzed for CD62L and JCV antibody index. Patient cohorts were grouped according to age, previous immune-suppression (IS), or JCV seropositivity and subsequently correlation between the two risk parameters 1) JCV index and 2) CD62L value was assessed. Results and conclusions: JCV seropositivity correlates with age, whereas CD62L values correlate with age and with duration of treatment. Interestingly, IS patients show a stronger correlation between CD62L values and treatment duration, suggesting CD62L as a first biological marker explaining why IS patients are at higher risk to develop PML over time. CD62L and JCV antibody indices only correlated in patients not previously IS (n = 226; p = 0.02). Risk stratification with the two markers alone set 43.37% at risk (JCV index N 0.9 and IS JCV + patients), or 3.74% (CD62L b 21.6). Synergy of the two markers reduced the percentage of patients at risk in our cohort (n = 273) to 2.18%. This data set suggests a statistical correlation between CD62L and JCV index, indicating a possible biological link between the two parameters. However, this link only exists without previous IS, which is in line with the observation that IS patients do not have higher JCV indices before developing PML. For patients with previous IS CD62L therefore seems favorable for risk stratification. For non-IS patients, the presented correlation and synergy between the two markers might have implications for the research of biological mechanisms leading to PML and can also help to further individualize risk stratification in long-term natalizumab patients. doi:10.1016/j.jneuroim.2014.08.067
Abstracts
Objective: Autoantibodies targeting aquaporin-4 (AQP4) are an important biomarker in neuromyelitis optica. Furthermore, autoantibodies to myelin oligodendrocyte glycoprotein (MOG) are detected in pediatric demyelinating disorders. We examined a cohort of adults with AQP4 antibody-negative neuromyelitis optica spectrum disorder (NMOSD) for antibodies to MOG. Methods: We performed a flow cytometry cell-based assay using live human embryonic kidney 293 cells that were lentivirus-transduced cells to express full-length surface MOG. Serum was tested in 23 AQP4 antibody-negative NMOSD patients with bilateral and/or recurrent optic neuritis (BON, n = 11), longitudinally extensive transverse myelitis (LETM, n = 10), and sequential BON and LETM (n = 2). Control cohorts included patients with clinically definite multiple sclerosis (MS, n = 76), as well as age matched healthy and other neurological disease controls (n = 52). Results: MOG antibodies were detected in 9/23 AQP4 antibodynegative NMOSD patients compared to 1/76 MS patients and 0/52 controls (P b 0.001). In all patients, the MOG antibodies were of the IgG rather than the IgM isotype. MOG antibodies were detected in 8/11 BON, 0/10 LETM, and 1/2 sequential BON and LETM patients. 6/9 MOG antibody-positive AQP4 antibody-negative patients had a relapsing course. MOG antibody-positive patients were more often female, of younger age at disease onset, and had a preceding viral prodrome compared to MOG antibody-negative patients (P values not significant). MOG antibody-positive patients had prominent bilateral optic disc swelling at presentation, and were more likely to have a rapid response to steroid therapy and relapse on steroid cessation than MOG antibody-negative patients (P = 0.034, P = 0.029, respectively). While 8/9 MOG antibody-positive patients had good follow-up visual acuity, one experienced sustained impairments in visual acuity and visual field testing. Furthermore, three patients had retinal nerve fiber layer atrophy on optical coherence tomography at follow-up, and one had residual spinal disability. Conclusions: MOG antibodies have a strong association with BON and may be a useful clinical biomarker. MOG antibody-associated BON is a relapsing disorder that is frequently steroid responsive and often steroid dependent. Failure of early recognition and institution of immunotherapy may be associated with sustained impairment. doi:10.1016/j.jneuroim.2014.08.065
113 Identification of candidate predictive protein biomarkers by M2 proteomics for clinical onset and treatment efficacy of multiple sclerosis Itay Raphael, Thomas G. Forsthuber The University of Texas at San Antonio, The University of Texas at San Antonio, San Antonio, United States Despite extensive research, multiple sclerosis (MS) remains a disease that lacks a definitive diagnostic test to predict imminent disease relapses. Thus, patients may undergo years of unnecessary treatments. Additionally, current treatments for MS can produce dramatically different outcomes in different individuals and therefore there is a critical need to develop biomarkers for treatment efficacy and resistance. We have recently developed a novel quantitative Microwave & Magnetic (M2) proteomics method to quantitatively measure changes in proteome expression over the course of experimental autoimmune encephalomyelitis (EAE), the standard murine animal model of MS. Our statistical analyses, including protein trajectories and receiver operating characteristic
(ROC) curves, indicate a strong correlation to EAE severity, and/or clinical-phase (i.e. time). Interestingly, M2 proteomics revealed characteristic CNS-specific protein expression waves prior to the onset of clinical symptoms. We are currently testing whether these characteristic protein expression waves allow us to predict the onset of clinical symptoms and forecast the severity of the disease. Furthermore, we have identified changes in the CNS proteome during EAE that correlate with the therapeutic efficacy of glucocorticoid treatment. Our studies will provide proof-of-principle for developing homologous human biomarkers that may be useful to predict disease onset, clinical severity and treatment efficacy. Finally, the changes in the CNS proteome detected by M2 proteomics may provide insights into key mechanisms that contribute to the disease pathology and may be useful to develop new therapeutic targets for MS. doi:10.1016/j.jneuroim.2014.08.066
495 JCV index and L-selectin for natalizumab-associated PML risk stratification Nicholas Schwab, Tilman Schneider-Hohendorf, Johanna Breuer, Anita Posevitz-Fejfar, Heinz Wiendl Neurology, University of Muenster, Muenster, Germany Background: Long-term treatment with and the presence of anti-JCV antibodies in serum are associated with the risk to develop-induced PML. Recent data suggest that the level of anti-JCV antibodies in serum (JCV antibody index N 0.9) and/or the lack of L-selectin (CD62L) on cryopreserved CD4+ T cells (%CD62L + cells of CD4+ T cells b 21.6) could be biomarkers for higher PML risk. Objectives: To compare and correlate PML risk stratification parameters during natalizumab therapy including the assessment of their relationship. Methods: Up to 1921 patients (number depending on data set completion) were analyzed for CD62L and JCV antibody index. Patient cohorts were grouped according to age, previous immune-suppression (IS), or JCV seropositivity and subsequently correlation between the two risk parameters 1) JCV index and 2) CD62L value was assessed. Results and conclusions: JCV seropositivity correlates with age, whereas CD62L values correlate with age and with duration of treatment. Interestingly, IS patients show a stronger correlation between CD62L values and treatment duration, suggesting CD62L as a first biological marker explaining why IS patients are at higher risk to develop PML over time. CD62L and JCV antibody indices only correlated in patients not previously IS (n = 226; p = 0.02). Risk stratification with the two markers alone set 43.37% at risk (JCV index N 0.9 and IS JCV + patients), or 3.74% (CD62L b 21.6). Synergy of the two markers reduced the percentage of patients at risk in our cohort (n = 273) to 2.18%. This data set suggests a statistical correlation between CD62L and JCV index, indicating a possible biological link between the two parameters. However, this link only exists without previous IS, which is in line with the observation that IS patients do not have higher JCV indices before developing PML. For patients with previous IS CD62L therefore seems favorable for risk stratification. For non-IS patients, the presented correlation and synergy between the two markers might have implications for the research of biological mechanisms leading to PML and can also help to further individualize risk stratification in long-term natalizumab patients. doi:10.1016/j.jneuroim.2014.08.067