Expression and targeting of transcription factor ATF5 in dog gliomas - PubMed (original) (raw)

. 2018 Mar;16(1):102-107.

doi: 10.1111/vco.12317. Epub 2017 May 8.

Affiliations

Expression and targeting of transcription factor ATF5 in dog gliomas

D York et al. Vet Comp Oncol. 2018 Mar.

Abstract

Background: Activating transcription factor 5 (ATF5) is a transcription factor that is highly expressed in undifferentiated neural progenitor/stem cells as well as a variety of human cancers including gliomas.

Aims: In this study, we examined the expression and localization of ATF5 protein in canine gliomas, and targeting of ATF5 function in canine glioma cell lines.

Materials and methods: Paraffin-embedded canine brain glioma tissue sections and western blots of tumours and glioma cells were immunoassayed with anti-ATF5 antibody. Viability of glioma cells was tested with a synthetic cell-penetrating ATF5 peptide (CP-d/n ATF5) ATF5 antagonist.

Results: ATF5 protein expression was in the nucleus and cytoplasm and was present in normal adult brain and tumour samples, with significantly higher expression in tumours as shown by western immunoblotting. CP-d/n ATF5 was found to decrease cell viability in canine glioma cell lines in vitro in a dose-dependent manner.

Conclusion: Similarities in expression of ATF5 in rodent, dog and human tumours, and cross species efficacy of the CP-d/n ATF5 peptide support the development of this ATF5-targeting approach as a novel and translational therapy in dog gliomas.

Keywords: astrocytoma; brain tumour; canine glioma cell lines; cell-penetrating peptide; d/n-ATF5; oligodendroglioma.

© 2017 John Wiley & Sons Ltd.

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Conflict of interest statement

Conflict of interest.

Columbia University, on behalf of inventors Dr. Angelastro awarded United States patents US 07888326 and US 08158420. Columbia University/University of California, Davis; Provisional patent application was filed, February 22, 2013 with U.S. Provisional Application Serial No. 61/768,390. Dr. Angelastro is a consultant/advisory board member for Sapience Therapeutics who hold technology licenses for ATF5 therapeutic targeting (patents held by Columbia University). Drs. DY, CDS, PJD, and NLC declare no potential conflict of interest.

Figures

Fig 1

Fig 1

Western immuno blot of ATF5 protein in canine normal brain, gliomas, and glioma cell lines J3TBg (JBg), SDT3G (SDT), and G06A (G06). Expression is seen in all samples (A), with higher expression seen generally in higher grade tumors compared to normal brain (B,C,D). NB= normal brain; A1,2,3,5=Grade II astrocytoma, A4=Grade III astrocytoma; G=glioblastoma/grade IV astrocytoma; O=Grade III oligodendroglioma, M1,4=Grade II oligoastrocytoma, M2,3=Grade III oligoastrocytoma. (** p=0.0037) (**** p<0.0001).

Fig 2

Fig 2

ATF5 fluorescent immunostaining of two canine glioblastoma samples (A–D and E–H). (A,E) ATF5 immunostaining, (C,G) DAPI nuclear staining, (B,F) ATF5/DAPI overlay, (D,H) ATF5 negative control (NC) consisting of non-relevant rabbit immunoglobulin. Scale bar = 20μm

Fig 3

Fig 3

ATF5 fluorescent immunostaining of peritumoral region from Fig 2A. (A) ATF5 immunostaining, (B) DAPI nuclear staining, (C) ATF5/DAPI overlay. Decreased cellularity and ATF5 immunostaining is apparent in the peritumoral tissue (P) compared to the tumor (T). Scale bar = 40μm

Fig 4

Fig 4

MTT cell viability dose response curves for canine glioma cell lines following exposure to CP-d/n-ATF5 peptide. Peptide effects were assessed in low serum (A,C) and standard serum (B,D) conditions. Values are means +/− standard deviations. (* significantly different from vehicle control, p<0.05)

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