Detection of IDH1 mutation in human gliomas: comparison of immunohistochemistry and sequencing

Abstract

Isocitrate dehydrogenase 1 (IDH1) mutations have recently been identified as early and frequent genetic alterations in astrocytomas, oligodendrogliomas, and oligoastrocytomas, as well as secondary glioblastomas, whereas primary glioblastomas very rarely contain IDH1 mutations. Furthermore, a specific monoclonal antibody, IMab-1, which recognizes IDH1-R132H-the most frequent IDH1 mutation-has been generated. IMab-1 has been reported to react with the IDH1-R132H protein, but not the wild-type IDH1 or the other IDH1 mutant proteins in Western-blot analysis. However, the importance of immunohistochemistry using IMab-1 has not yet been elucidated. In this study, we compared the findings from IMab-1 immunohistochemistry and direct DNA sequencing using 49 glioma samples. IMab-1 detected 12 out of 49 cases; however, only nine cases were found to be IDH1-R132H by direct DNA sequencing because of a small population of IDH1-R132H mutation-possessing tumor cells, indicating that IMab-1 immunohistochemistry is useful for detecting IDH1-R132H. We conducted immunohistochemical detection in 52 cases of grade III astrocytomas. The median time to progression (TTP) was significantly longer in the cases with the IDH1 mutation (86.7 months) compared to the cases without the IDH1 mutation (wild type, 10.4 months) (p < 0.01). In conclusion, the anti-IDH1-R132H-specific monoclonal antibody IMab-1 is very useful for detecting IDH1-R132H in immunohistochemistry, and predicting the time to progression in grade III anaplastic astrocytomas. Therefore, IMab-1 is likely to be useful for the diagnosis of mutation-bearing gliomas and for determining the treatment strategy of grade III gliomas.

Diffusely infiltrating astrocytomas: pathology, molecular mechanisms and markers

“…The commercially available antibodies for IDH1 R132H are highly specific and strongly stain the cytoplasm of IDH1 R132H harbouring tumour cells but not non-neoplastic cells [23]. Both high accuracy and sensitivity of immunodetection, even in a mixed cell population, have been demonstrated [138]. Monoclonal antibodies against other types of mutant IDH1 or IDH2, some of which are specific for more than one type of mutation, are now also available, technically enabling the detection of almost all types of IDH1/2 mutations except IDH1 R132C [63].…”

Can Diffusion Tensor Imaging Noninvasively Detect IDH1 Gene Mutations in Astrogliomas? A Retrospective Study of 112 Cases

“…IDH1 R132H protein expression was determined in paraffin-embedded tumor specimens after surgery by use of anti-mIDH1R132H antibody (internal clone H09; Dianova, Hamburg, Germany). The expression of IDH1 R132H was determined by use of the methods of Takano et al 17 Cases with Ն10% positive cells were rated positive, and cases with Ͻ10% positive cells were rated negative. There were 25 (69.44%), 12 (52.17%), and 10 (18.87%) cases showing positive results in grades II, III, and IV, respectively.…”

Evaluation of the Role of Mutant Idh1, Egfr and Bcl2 Protein Expression in Distinguishing Diffuse Astrocytoma From Astrocytosis

“…A controversial result was given by Takano et al, 2011 [27] who reported that; IDH1 expression in diffuse astrocytoma grade II were 100%; these results were different from the current results that may be due to different used antibody clone; where they used clone MT 3.1 (Neomarker, Westinghouse, Fremont, CA, USA); while in the present study anti IDH1 rabbit monoclonal antibody clone H09 (Cell signaling Technology, Dinova, Hamburg, Germany) was used. Also their study involved only four cases; this may be other cause.…”