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In situ Protein Detection for Companion Diagnostics.

Gremel G, Grannas K, Sutton LA, Pontén F, Zieba A - Front Oncol (2013)

Bottom Line: The emergence of targeted therapies for cancer has created a need for the development of companion diagnostic tests.We also explore the possibility of using IHC to detect specific protein mutations in addition to DNA-based tests.Finally, we review alternative protein binders and proximity ligation assays and discuss their potential to facilitate the development of novel, targeted therapies against cancer.

View Article: PubMed Central - PubMed

Affiliation: Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University , Uppsala , Sweden.

ABSTRACT
The emergence of targeted therapies for cancer has created a need for the development of companion diagnostic tests. Assays developed in recent years are aimed at determining both the effectiveness and safety of specific drugs for a defined group of patients, thus, enabling the more efficient design of clinical trials and also supporting physicians when making treatment-related decisions. Immunohistochemistry (IHC) is a widely accepted method for protein expression analyses in human tissues. Immunohistochemical assays, used to localize and quantitate relative protein expression levels within a morphological context, are frequently used as companion diagnostics during clinical trials and also following drug approval. Herein, we describe established immunochemistry-based methods and their application in routine diagnostics. We also explore the possibility of using IHC to detect specific protein mutations in addition to DNA-based tests. Finally, we review alternative protein binders and proximity ligation assays and discuss their potential to facilitate the development of novel, targeted therapies against cancer.

No MeSH data available.


Related in: MedlinePlus

BRAFV600E mutation-specific antibody staining. Immunohistochemical staining examples of the BRAFV600E mutation-specific antibody VE1 are presented for a BRAFV600E-positive and a BRAFV600E-negative case of melanoma, colon cancer, and thyroid cancer, respectively. BRAFV600E-positive staining is generally detected as a granular, cytoplasmic signal that can easily be distinguished from BRAFV600E-negative cases. In the presented example images, 3,3′-Diaminobenzidine (DAB) was used as a chromogen.
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Figure 1: BRAFV600E mutation-specific antibody staining. Immunohistochemical staining examples of the BRAFV600E mutation-specific antibody VE1 are presented for a BRAFV600E-positive and a BRAFV600E-negative case of melanoma, colon cancer, and thyroid cancer, respectively. BRAFV600E-positive staining is generally detected as a granular, cytoplasmic signal that can easily be distinguished from BRAFV600E-negative cases. In the presented example images, 3,3′-Diaminobenzidine (DAB) was used as a chromogen.

Mentions: The v-raf murine sarcoma viral oncogene homolog B1 (BRAF) represents an outstanding target for the development of a mutation-specific antibody. Mutations of the associated gene occur in a range of human malignancies including cutaneous melanoma, colorectal cancer, NSCLC, papillary thyroid cancer, and hairy-cell leukemia (57–59). By far, the most common BRAF mutation results in the substitution of valine for glutamic acid at position 600 (V600E), leading to the constitutive activation of the protein’s kinase domain. In human cutaneous melanoma, mutated BRAF has been detected in 40–50% of cases, with up 90% of these alterations concerning the V-E substitution at codon 600 (60). Vemurafenib and dabrafenib are two potent small molecule inhibitor drugs that specifically target BRAF V600E and have demonstrated remarkable response rates in metastatic melanoma patients (61, 62). Mutational testing of the patient tumor material is required before commencement of treatment and is to date commonly based on the detection of genomic alterations. As an addition to DNA testing, Capper et al. recently proposed a mutation-specific antibody for the detection of BRAF V600E in FFPE tissue specimens by means of IHC (Figure 1) (63). The results obtained from using this antibody to determine the BRAF mutational status in melanoma and thyroid cancer samples were identical to those achieved following DNA sequencing-based profiling. These results have since been substantiated by numerous studies with similarly high levels of specificity and sensitivity (up to 100%, respectively) (64–68).


In situ Protein Detection for Companion Diagnostics.

Gremel G, Grannas K, Sutton LA, Pontén F, Zieba A - Front Oncol (2013)

BRAFV600E mutation-specific antibody staining. Immunohistochemical staining examples of the BRAFV600E mutation-specific antibody VE1 are presented for a BRAFV600E-positive and a BRAFV600E-negative case of melanoma, colon cancer, and thyroid cancer, respectively. BRAFV600E-positive staining is generally detected as a granular, cytoplasmic signal that can easily be distinguished from BRAFV600E-negative cases. In the presented example images, 3,3′-Diaminobenzidine (DAB) was used as a chromogen.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3814083&req=5

Figure 1: BRAFV600E mutation-specific antibody staining. Immunohistochemical staining examples of the BRAFV600E mutation-specific antibody VE1 are presented for a BRAFV600E-positive and a BRAFV600E-negative case of melanoma, colon cancer, and thyroid cancer, respectively. BRAFV600E-positive staining is generally detected as a granular, cytoplasmic signal that can easily be distinguished from BRAFV600E-negative cases. In the presented example images, 3,3′-Diaminobenzidine (DAB) was used as a chromogen.
Mentions: The v-raf murine sarcoma viral oncogene homolog B1 (BRAF) represents an outstanding target for the development of a mutation-specific antibody. Mutations of the associated gene occur in a range of human malignancies including cutaneous melanoma, colorectal cancer, NSCLC, papillary thyroid cancer, and hairy-cell leukemia (57–59). By far, the most common BRAF mutation results in the substitution of valine for glutamic acid at position 600 (V600E), leading to the constitutive activation of the protein’s kinase domain. In human cutaneous melanoma, mutated BRAF has been detected in 40–50% of cases, with up 90% of these alterations concerning the V-E substitution at codon 600 (60). Vemurafenib and dabrafenib are two potent small molecule inhibitor drugs that specifically target BRAF V600E and have demonstrated remarkable response rates in metastatic melanoma patients (61, 62). Mutational testing of the patient tumor material is required before commencement of treatment and is to date commonly based on the detection of genomic alterations. As an addition to DNA testing, Capper et al. recently proposed a mutation-specific antibody for the detection of BRAF V600E in FFPE tissue specimens by means of IHC (Figure 1) (63). The results obtained from using this antibody to determine the BRAF mutational status in melanoma and thyroid cancer samples were identical to those achieved following DNA sequencing-based profiling. These results have since been substantiated by numerous studies with similarly high levels of specificity and sensitivity (up to 100%, respectively) (64–68).

Bottom Line: The emergence of targeted therapies for cancer has created a need for the development of companion diagnostic tests.We also explore the possibility of using IHC to detect specific protein mutations in addition to DNA-based tests.Finally, we review alternative protein binders and proximity ligation assays and discuss their potential to facilitate the development of novel, targeted therapies against cancer.

View Article: PubMed Central - PubMed

Affiliation: Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University , Uppsala , Sweden.

ABSTRACT
The emergence of targeted therapies for cancer has created a need for the development of companion diagnostic tests. Assays developed in recent years are aimed at determining both the effectiveness and safety of specific drugs for a defined group of patients, thus, enabling the more efficient design of clinical trials and also supporting physicians when making treatment-related decisions. Immunohistochemistry (IHC) is a widely accepted method for protein expression analyses in human tissues. Immunohistochemical assays, used to localize and quantitate relative protein expression levels within a morphological context, are frequently used as companion diagnostics during clinical trials and also following drug approval. Herein, we describe established immunochemistry-based methods and their application in routine diagnostics. We also explore the possibility of using IHC to detect specific protein mutations in addition to DNA-based tests. Finally, we review alternative protein binders and proximity ligation assays and discuss their potential to facilitate the development of novel, targeted therapies against cancer.

No MeSH data available.


Related in: MedlinePlus