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Equivalent Gene Expression Profiles between Glatopa™ and Copaxone®.

D'Alessandro JS, Duffner J, Pradines J, Capila I, Garofalo K, Kaundinya G, Greenberg BM, Kantor D, Ganguly TC - PLoS ONE (2015)

Bottom Line: Comparative analysis was also performed on sets of transcripts relevant to T-cell biology and antigen presentation, among others that are known to be modulated by glatiramer acetate.No statistically significant differences were observed between Copaxone and Glatopa in the expression levels (magnitude and direction) of these glatiramer acetate-regulated genes.In conclusion, multiple methods consistently supported equivalent gene expression profiles between Copaxone and Glatopa.

View Article: PubMed Central - PubMed

Affiliation: Momenta Pharmaceuticals, Inc., Cambridge, MA, United States of America.

ABSTRACT
Glatopa™ is a generic glatiramer acetate recently approved for the treatment of patients with relapsing forms of multiple sclerosis. Gene expression profiling was performed as a means to evaluate equivalence of Glatopa and Copaxone®. Microarray analysis containing 39,429 unique probes across the entire genome was performed in murine glatiramer acetate--responsive Th2-polarized T cells, a test system highly relevant to the biology of glatiramer acetate. A closely related but nonequivalent glatiramoid molecule was used as a control to establish assay sensitivity. Multiple probe-level (Student's t-test) and sample-level (principal component analysis, multidimensional scaling, and hierarchical clustering) statistical analyses were utilized to look for differences in gene expression induced by the test articles. The analyses were conducted across all genes measured, as well as across a subset of genes that were shown to be modulated by Copaxone. The following observations were made across multiple statistical analyses: the expression of numerous genes was significantly changed by treatment with Copaxone when compared against media-only control; gene expression profiles induced by Copaxone and Glatopa were not significantly different; and gene expression profiles induced by Copaxone and the nonequivalent glatiramoid were significantly different, underscoring the sensitivity of the test system and the multiple analysis methods. Comparative analysis was also performed on sets of transcripts relevant to T-cell biology and antigen presentation, among others that are known to be modulated by glatiramer acetate. No statistically significant differences were observed between Copaxone and Glatopa in the expression levels (magnitude and direction) of these glatiramer acetate-regulated genes. In conclusion, multiple methods consistently supported equivalent gene expression profiles between Copaxone and Glatopa.

No MeSH data available.


Related in: MedlinePlus

Characterization of the murine Th2-polarized T cells.Cytokine profile and dose response. A: Cytokine profile of the conditioned medium from the Th2-455 cell line after 24 hours of treatment with Copaxone at a single concentration of 20 μg/mL, demonstrating Th2 polarization. B: Copaxone dose response at 24 hours of a single cytokine, IL-4. IFN-γ; interferon gamma; KC, keratinocyte chemoattractant; IL, interleukin; TGF-β1, transforming growth factor-beta 1; Th, T-helper.
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pone.0140299.g002: Characterization of the murine Th2-polarized T cells.Cytokine profile and dose response. A: Cytokine profile of the conditioned medium from the Th2-455 cell line after 24 hours of treatment with Copaxone at a single concentration of 20 μg/mL, demonstrating Th2 polarization. B: Copaxone dose response at 24 hours of a single cytokine, IL-4. IFN-γ; interferon gamma; KC, keratinocyte chemoattractant; IL, interleukin; TGF-β1, transforming growth factor-beta 1; Th, T-helper.

Mentions: The phenotype of the GA-responsive murine Th2 T cells was confirmed by single and multiplexed ELISAs for Th1 and Th2 cytokines. These cells secreted low levels of the Th1 cytokines (IL-1β, TNF-α, and IFN-γ), whereas the Th2 cytokines (IL-4, IL-5, IL-10, and IL-13) were released at high concentrations (Fig 2A). Copaxone produced a dose-dependent increase in IL-4 release from these Th2-polarized T cells (Fig 2B); a similar dose effect was seen with the other cytokines (data not shown). Whole-genome gene expression of multiple samples of Copaxone, in 17 samples from 9 different lots, were compared with those of 7 samples of media-only controls. The number of probes that were significantly different was analyzed using univariate analysis (Student’s t-test). The results of univariate analysis (Table 1) indicated that of the 39,429 probes evaluated at the probe level by pairwise comparison, 9815 probes were significantly different at a P-value threshold of 0.05 (before controlling for false positives) when comparing Copaxone with media-only control. Evaluation of the data using FDR (threshold q = 0.05) or the more conservative FWER with Bonferroni correction test indicated that 6869 and 1080 probes, respectively, were significantly different in Copaxone than in the media-only control. Permutation control analysis on this data set indicated that these differences were not attributed to random sampling of the data, as indicated by the significance (P < 0.0005) of observing the given number of probes compared with the hypothesis of no difference between groups.


Equivalent Gene Expression Profiles between Glatopa™ and Copaxone®.

D'Alessandro JS, Duffner J, Pradines J, Capila I, Garofalo K, Kaundinya G, Greenberg BM, Kantor D, Ganguly TC - PLoS ONE (2015)

Characterization of the murine Th2-polarized T cells.Cytokine profile and dose response. A: Cytokine profile of the conditioned medium from the Th2-455 cell line after 24 hours of treatment with Copaxone at a single concentration of 20 μg/mL, demonstrating Th2 polarization. B: Copaxone dose response at 24 hours of a single cytokine, IL-4. IFN-γ; interferon gamma; KC, keratinocyte chemoattractant; IL, interleukin; TGF-β1, transforming growth factor-beta 1; Th, T-helper.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608686&req=5

pone.0140299.g002: Characterization of the murine Th2-polarized T cells.Cytokine profile and dose response. A: Cytokine profile of the conditioned medium from the Th2-455 cell line after 24 hours of treatment with Copaxone at a single concentration of 20 μg/mL, demonstrating Th2 polarization. B: Copaxone dose response at 24 hours of a single cytokine, IL-4. IFN-γ; interferon gamma; KC, keratinocyte chemoattractant; IL, interleukin; TGF-β1, transforming growth factor-beta 1; Th, T-helper.
Mentions: The phenotype of the GA-responsive murine Th2 T cells was confirmed by single and multiplexed ELISAs for Th1 and Th2 cytokines. These cells secreted low levels of the Th1 cytokines (IL-1β, TNF-α, and IFN-γ), whereas the Th2 cytokines (IL-4, IL-5, IL-10, and IL-13) were released at high concentrations (Fig 2A). Copaxone produced a dose-dependent increase in IL-4 release from these Th2-polarized T cells (Fig 2B); a similar dose effect was seen with the other cytokines (data not shown). Whole-genome gene expression of multiple samples of Copaxone, in 17 samples from 9 different lots, were compared with those of 7 samples of media-only controls. The number of probes that were significantly different was analyzed using univariate analysis (Student’s t-test). The results of univariate analysis (Table 1) indicated that of the 39,429 probes evaluated at the probe level by pairwise comparison, 9815 probes were significantly different at a P-value threshold of 0.05 (before controlling for false positives) when comparing Copaxone with media-only control. Evaluation of the data using FDR (threshold q = 0.05) or the more conservative FWER with Bonferroni correction test indicated that 6869 and 1080 probes, respectively, were significantly different in Copaxone than in the media-only control. Permutation control analysis on this data set indicated that these differences were not attributed to random sampling of the data, as indicated by the significance (P < 0.0005) of observing the given number of probes compared with the hypothesis of no difference between groups.

Bottom Line: Comparative analysis was also performed on sets of transcripts relevant to T-cell biology and antigen presentation, among others that are known to be modulated by glatiramer acetate.No statistically significant differences were observed between Copaxone and Glatopa in the expression levels (magnitude and direction) of these glatiramer acetate-regulated genes.In conclusion, multiple methods consistently supported equivalent gene expression profiles between Copaxone and Glatopa.

View Article: PubMed Central - PubMed

Affiliation: Momenta Pharmaceuticals, Inc., Cambridge, MA, United States of America.

ABSTRACT
Glatopa™ is a generic glatiramer acetate recently approved for the treatment of patients with relapsing forms of multiple sclerosis. Gene expression profiling was performed as a means to evaluate equivalence of Glatopa and Copaxone®. Microarray analysis containing 39,429 unique probes across the entire genome was performed in murine glatiramer acetate--responsive Th2-polarized T cells, a test system highly relevant to the biology of glatiramer acetate. A closely related but nonequivalent glatiramoid molecule was used as a control to establish assay sensitivity. Multiple probe-level (Student's t-test) and sample-level (principal component analysis, multidimensional scaling, and hierarchical clustering) statistical analyses were utilized to look for differences in gene expression induced by the test articles. The analyses were conducted across all genes measured, as well as across a subset of genes that were shown to be modulated by Copaxone. The following observations were made across multiple statistical analyses: the expression of numerous genes was significantly changed by treatment with Copaxone when compared against media-only control; gene expression profiles induced by Copaxone and Glatopa were not significantly different; and gene expression profiles induced by Copaxone and the nonequivalent glatiramoid were significantly different, underscoring the sensitivity of the test system and the multiple analysis methods. Comparative analysis was also performed on sets of transcripts relevant to T-cell biology and antigen presentation, among others that are known to be modulated by glatiramer acetate. No statistically significant differences were observed between Copaxone and Glatopa in the expression levels (magnitude and direction) of these glatiramer acetate-regulated genes. In conclusion, multiple methods consistently supported equivalent gene expression profiles between Copaxone and Glatopa.

No MeSH data available.


Related in: MedlinePlus