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Quality assessment of cellular therapies: the emerging role of molecular assays.

Stroncek DF, Jin P, Ren J, Feng J, Castiello L, Civini S, Wang E, Marincola FM, Sabatino M - Korean J Hematol (2010)

Bottom Line: These therapies are becoming more effective and are being used more frequently, but they are also becoming more complex.Products are evaluated at critical steps in the manufacturing process and at the end of production prior to the release of the product for clinical use.We and others have found that global gene and microRNA expression analysis is useful for comparability testing and will likely be useful for potency, stability and consistency testing.

View Article: PubMed Central - PubMed

Affiliation: Cellular Therapy and Immunogenetics Sections, Department of Transfusion Medicine, Clinical Center, NIH, Bethesda, MD, USA.

ABSTRACT
Cellular therapies are becoming increasingly important in treating cancer, hematologic malignancies, autoimmune disorders, and damaged tissue. These therapies are becoming more effective and are being used more frequently, but they are also becoming more complex. As a result, quality testing is becoming an increasingly important part of cellular therapy. Cellular therapies should be tested at several points during their production. The starting material, intermediate products and the final product are usually analyzed. Products are evaluated at critical steps in the manufacturing process and at the end of production prior to the release of the product for clinical use. In addition, the donor of the starting biologic material is usually evaluated. The testing of cellular therapies for stability, consistency, comparability and potency is especially challenging. We and others have found that global gene and microRNA expression analysis is useful for comparability testing and will likely be useful for potency, stability and consistency testing. Several examples of the use of gene expression analysis for assessing cellular therapies are presented.

No MeSH data available.


Related in: MedlinePlus

Global gene expression analysis separated immature dentritic cells (iDCs) and mature dentritic cells (mDCs) by donor rather than maturation cocktail. Three sets of iDCs were prepared from six healthy subjects (donors 1 through 6) by culturing peripheral blood monocytes with IL-4 and GM-CSF for 3 days. The iDC samples of each subject were matured by culture with one of three maturation cocktails: LPS plus IFN-γ(C1); LPS, IFN-γ plus IL-1β(C2); and LPS, IFN-γ, IL-1β plus TNF-α(C3). The 18 iDC and 18 mDC samples were analyzed by global gene expression profiling with more than 36,000 oligonucleotide probes and the results were analyzed by unsupervised hierarchical clustering analysis. The iDC and mDC samples were in two separate clusters. Within these two clusters, the samples grouped according to subject. ANOVA analysis revealed that expression of 9,590 genes were differentially expressed among donors, but only 13 genes were differentially expressed among differentiation protocols [33].
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Figure 1: Global gene expression analysis separated immature dentritic cells (iDCs) and mature dentritic cells (mDCs) by donor rather than maturation cocktail. Three sets of iDCs were prepared from six healthy subjects (donors 1 through 6) by culturing peripheral blood monocytes with IL-4 and GM-CSF for 3 days. The iDC samples of each subject were matured by culture with one of three maturation cocktails: LPS plus IFN-γ(C1); LPS, IFN-γ plus IL-1β(C2); and LPS, IFN-γ, IL-1β plus TNF-α(C3). The 18 iDC and 18 mDC samples were analyzed by global gene expression profiling with more than 36,000 oligonucleotide probes and the results were analyzed by unsupervised hierarchical clustering analysis. The iDC and mDC samples were in two separate clusters. Within these two clusters, the samples grouped according to subject. ANOVA analysis revealed that expression of 9,590 genes were differentially expressed among donors, but only 13 genes were differentially expressed among differentiation protocols [33].

Mentions: Global gene expression analysis may be an excellent tool for assessing the consistency of cellular therapies. In the study described above, in which we evaluated mDCs produced with three different maturation cocktails (LPS plus IFN-γ; LPS, IFN-γ plus IL-1β; and LPS, IFN-γ, IL-1β plus TNF-α) from six healthy subjects, the DCs clustered according to donor rather than maturation protocol (Fig. 1). This result suggests that global gene expression profiling can distinguish differences due to donor variability. The ability of gene expression profiling to detect minor differences between healthy subjects will likely make this an excellent assay for consistency testing.


Quality assessment of cellular therapies: the emerging role of molecular assays.

Stroncek DF, Jin P, Ren J, Feng J, Castiello L, Civini S, Wang E, Marincola FM, Sabatino M - Korean J Hematol (2010)

Global gene expression analysis separated immature dentritic cells (iDCs) and mature dentritic cells (mDCs) by donor rather than maturation cocktail. Three sets of iDCs were prepared from six healthy subjects (donors 1 through 6) by culturing peripheral blood monocytes with IL-4 and GM-CSF for 3 days. The iDC samples of each subject were matured by culture with one of three maturation cocktails: LPS plus IFN-γ(C1); LPS, IFN-γ plus IL-1β(C2); and LPS, IFN-γ, IL-1β plus TNF-α(C3). The 18 iDC and 18 mDC samples were analyzed by global gene expression profiling with more than 36,000 oligonucleotide probes and the results were analyzed by unsupervised hierarchical clustering analysis. The iDC and mDC samples were in two separate clusters. Within these two clusters, the samples grouped according to subject. ANOVA analysis revealed that expression of 9,590 genes were differentially expressed among donors, but only 13 genes were differentially expressed among differentiation protocols [33].
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2983004&req=5

Figure 1: Global gene expression analysis separated immature dentritic cells (iDCs) and mature dentritic cells (mDCs) by donor rather than maturation cocktail. Three sets of iDCs were prepared from six healthy subjects (donors 1 through 6) by culturing peripheral blood monocytes with IL-4 and GM-CSF for 3 days. The iDC samples of each subject were matured by culture with one of three maturation cocktails: LPS plus IFN-γ(C1); LPS, IFN-γ plus IL-1β(C2); and LPS, IFN-γ, IL-1β plus TNF-α(C3). The 18 iDC and 18 mDC samples were analyzed by global gene expression profiling with more than 36,000 oligonucleotide probes and the results were analyzed by unsupervised hierarchical clustering analysis. The iDC and mDC samples were in two separate clusters. Within these two clusters, the samples grouped according to subject. ANOVA analysis revealed that expression of 9,590 genes were differentially expressed among donors, but only 13 genes were differentially expressed among differentiation protocols [33].
Mentions: Global gene expression analysis may be an excellent tool for assessing the consistency of cellular therapies. In the study described above, in which we evaluated mDCs produced with three different maturation cocktails (LPS plus IFN-γ; LPS, IFN-γ plus IL-1β; and LPS, IFN-γ, IL-1β plus TNF-α) from six healthy subjects, the DCs clustered according to donor rather than maturation protocol (Fig. 1). This result suggests that global gene expression profiling can distinguish differences due to donor variability. The ability of gene expression profiling to detect minor differences between healthy subjects will likely make this an excellent assay for consistency testing.

Bottom Line: These therapies are becoming more effective and are being used more frequently, but they are also becoming more complex.Products are evaluated at critical steps in the manufacturing process and at the end of production prior to the release of the product for clinical use.We and others have found that global gene and microRNA expression analysis is useful for comparability testing and will likely be useful for potency, stability and consistency testing.

View Article: PubMed Central - PubMed

Affiliation: Cellular Therapy and Immunogenetics Sections, Department of Transfusion Medicine, Clinical Center, NIH, Bethesda, MD, USA.

ABSTRACT
Cellular therapies are becoming increasingly important in treating cancer, hematologic malignancies, autoimmune disorders, and damaged tissue. These therapies are becoming more effective and are being used more frequently, but they are also becoming more complex. As a result, quality testing is becoming an increasingly important part of cellular therapy. Cellular therapies should be tested at several points during their production. The starting material, intermediate products and the final product are usually analyzed. Products are evaluated at critical steps in the manufacturing process and at the end of production prior to the release of the product for clinical use. In addition, the donor of the starting biologic material is usually evaluated. The testing of cellular therapies for stability, consistency, comparability and potency is especially challenging. We and others have found that global gene and microRNA expression analysis is useful for comparability testing and will likely be useful for potency, stability and consistency testing. Several examples of the use of gene expression analysis for assessing cellular therapies are presented.

No MeSH data available.


Related in: MedlinePlus