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Imaging HDACs In Vivo: Cross-Validation of the [11C]Martinostat Radioligand in the Pig Brain

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Abstract

Purpose

With the emerging knowledge about the impact of epigenetic alterations on behavior and brain disorders, the ability to measure epigenetic alterations in brain tissue in vivo has become critically important. We present the first in vivo/in vitro cross-validation of the novel positron emission tomography (PET) radioligand [11C]Martinostat in the pig brain with regard to its ability to measure histone deacetylase 1–3 (HDAC1–3) levels in vivo.

Procedures

Nine female Danish landrace pigs underwent 121-min dynamic PET scans with [11C]Martinostat. We quantified [11C]Martinostat uptake using both a simple ratio method and kinetic models with arterial input function. By the end of the scan, the animals were euthanized and the brains were extracted. We measured HDAC1–3 protein levels in frontal cortex, cerebellum vermis, and hippocampus and compared the protein levels and regional outcome values to the [11C]Martinostat PET quantification.

Results

[11C]Martinostat distributed widely across brain regions, with the highest uptake in the cerebellum vermis and the lowest in the olfactory bulbs. Based on the Akaike information criterion, the quantification was most reliably performed by Ichise MA1 kinetic modeling, but since the radioligand displayed very slow kinetics, we also calculated standard uptake value (SUV) ratios which correlated well with VT. The western blots revealed higher brain tissue protein levels of HDAC1/2 compared to HDAC3, and HDAC1 and HDAC2 levels were highly correlated in all three investigated brain regions. The in vivo SUV ratio measure correlated well with the in vitro HDAC1–3 levels, whereas no correlation was found between VT values and HDAC levels.

Conclusions

We found good correlation between in vivo measured SUV ratios and in vitro measures of HDAC 1–3 proteins, supporting that [11C]Martinostat provides a good in vivo measure of the cerebral HDAC1–3 protein levels.

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References

  1. Abel T, Zukin S (2008) Epigenetic targets of HDAC inhibitition in neurodegenerative and psychiartic disorders. Curr Opin Pharmacol 8:57–64

    Article  CAS  Google Scholar 

  2. Penney J, Tsai L-H (2014) Histone deacetylases in memory and cognition. Neuroscience 7:3–10

    Google Scholar 

  3. Gilbert TM, Zürcher NR, Wu CJ, Bhanot A, Hightower BG, Kim M, Albrecht DS, Wey HY, Schroeder FA, Rodriguez-Thompson A, Morin TM, Hart KL, Pellegrini AM, Riley MM, Wang C, Stufflebeam SM, Haggarty SJ, Holt DJ, Loggia ML, Perlis RH, Brown HE, Roffman JL, Hooker JM (2018) PET neuroimaging reveals histone deacetylase dysregulation in schizophrenia. J Clin Invest 129:364–372. https://doi.org/10.1172/JCI123743

    Article  PubMed  PubMed Central  Google Scholar 

  4. Wang C, Schroeder FA, Wey HY, Borra R, Wagner FF, Reis S, Kim SW, Holson EB, Haggarty SJ, Hooker JM (2014) In vivo imaging of histone deacetylases (HDACs) in the central nervous system and major peripheral organs. J Med Chem 57:7999–8009

    Article  CAS  Google Scholar 

  5. Wey H-Y, Wang C, Schroeder FA, Schroeder FA, Logan J, Price JC, Hooker JM (2015) Kinetic analysis and quantification of [11C]Martinostat for in vivo HDAC imaging of the brain. ACS Chem Neurosci 6:708–715

    Article  CAS  Google Scholar 

  6. Schroeder FA, Wang C, Van De Bittner GC et al (2014) PET imaging demonstrates histone deacetylase target engagement and clarifies brain penetrance of known and novel small molecule inhibitors in rat. ACS Chem Neurosci 5:1055–1062

    Article  CAS  Google Scholar 

  7. Wey H-Y, Gilbert TM, Zurcher NR et al (2016) Insights into neuroepigenetics through human histone deacetylase PET imaging. Sci Transl Med 8:351ra106. https://doi.org/10.1126/scitranslmed.aaf7551

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Gillings N (2009) A restricted access material for rapid analysis of [11C]-labeled radiopharmaceuticals and their metabolites in plasma. Nucl Med Biol 36:961–965

    Article  CAS  Google Scholar 

  9. Hong IK, Chung ST, Kim HK et al (2007) Ultra fast symmetry and SIMD-based projection-backprojection (SSP) algorithm for 3-D PET image reconstruction. IEEE Trans Med Imaging 6:789–803

    Article  Google Scholar 

  10. Sureau FC, Reader AJ, Comtat C, Leroy C, Ribeiro MJ, Buvat I, Trebossen R (2008) Impact of image-space resolution modeling for studies with the high-resolution research tomograph. J Nucl Med 49:1000–1008

    Article  Google Scholar 

  11. Keller SH, Svarer C, Sibomana M (2013) Attenuation correction for the HRRT PET-scanner using transmission scatter correction and total variation regularization. IEEE Trans Med Imaging 32:1611–1621

    Article  Google Scholar 

  12. Villadsen J, Hansen HD, Jørgensen LM et al (2017) Automatic delineation of brain regions on MRI and PET images from the pig. J Neurosci Methods 294:51–58

    Article  Google Scholar 

  13. Saikali S, Meurice P, Sauleau P, Eliat PA, Bellaud P, Randuineau G, Vérin M, Malbert CH (2010) A three-dimensional digital segmented and deformable brain atlas of the domestic pig. J Neurosci Methods 192:102–109

    Article  Google Scholar 

  14. Cunningham VJ, E a R, Slifstein M et al (2010) Measuring drug occupancy in the absence of a reference region: the Lassen plot re-visited. J Cereb Blood Flow Metab 30:46–50

    Article  Google Scholar 

  15. Seto E, Yoshida M (2014) Erasers of histone acetylation: the histone deacetylase enzymes. Cold Spring Harb Perspect Biol 6. https://doi.org/10.1101/cshperspect.a018713

    Article  Google Scholar 

  16. Anderson KW, Chen J, Wang M, Mast N, Pikuleva IA, Turko IV (2015) Quantification of histone deacetylase isoforms in human frontal cortex, human retina, and mouse brain. PLoS One 10:e0126592. https://doi.org/10.1371/journal.pone.0126592

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Wang Y, Zhang YL, Hennig K, Gale JP, Hong Y, Cha A, Riley M, Wagner F, Haggarty SJ, Holson E, Hooker J (2013) Class I HDAC imaging using [3H]CI-994 autoradiography. Epigenetics 8:756–764

    Article  CAS  Google Scholar 

  18. Chen YT, Zang XF, Pan J, Zhu XL, Chen F, Chen ZB, Xu Y (2012) Expression patterns of histone deacetylases in experimental stroke and potential targets for neuroprotection. Clin Exp Pharmacol Physiol 39:751–758

    Article  Google Scholar 

  19. Gu Y, Yang P, Shao Q et al (2013) Investigation of the expression patterns and correlation of DNA methyltransferases and class I histone deacetylases in ovarian cancer tissues. Oncol Lett 5:452–458

    Article  CAS  Google Scholar 

  20. Quint K, Agaimy A, Di Fazio P et al (2011) Clinical significance of histone deacetylases 1, 2, 3, and 7: HDAC2 is an independent predictor of survival in HCC. Virchows Arch 459:129–139

    Article  CAS  Google Scholar 

  21. Jurkin J, Zupkovitz G, Lagger S, Grausenburger R, Hagelkruys A, Kenner L, Seiser C (2011) Distinct and redundant functions of histone deacetylases HDAC1 and HDAC2 in proliferation and tumorigenesis. Cell Cycle 10:406–412

    Article  CAS  Google Scholar 

  22. Wilting RH, Yanover E, Heideman MR, Jacobs H, Horner J, van der Torre J, DePinho RA, Dannenberg JH (2010) Overlapping functions of Hdac1 and Hdac2 in cell cycle regulation and haematopoiesis. EMBO J 29:2586–2597

    Article  CAS  Google Scholar 

  23. Somanath P, Herndon Klein R, Knoepfler PS (2017) CRISPR-mediated HDAC2 disruption identifies two distinct classes of target genes in human cells. PLoS One 12:1–21

    Article  Google Scholar 

  24. Joksimovic SM, Osuru HP, Oklopcic A et al (2018) Histone deacetylase inhibitor entinostat (MS-275) restores anesthesia-induced alteration of inhibitory synaptic transmission in the developing rat hippocampus. Mol Neurobiol 55:22–228

    Article  Google Scholar 

  25. Gregoretti IV, Lee YM, Goodson HV (2004) Molecular evolution of the histone deacetylase family: functional implications of phylogenetic analysis. J Mol Biol 338:17–31

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to give thanks to C.A. Olsen for kindly providing recombinant HDAC proteins and the staff of Department of Experimental Medicine at University of Copenhagen for excellent assistance.

Funding

This study was funded by The Lundbeck Foundation (grant no. R180-2014-3171 and R233-2016-3416).

Author information

Authors and Affiliations

  1. Neurobiology Research Unit and Center for NeuroPharm, Copenhagen University Hospital Rigshospitalet, 9 Blegdamsvej, 2100, Copenhagen O, Denmark

    L. L. Donovan, J. H. Magnussen, A. Dyssegaard, G. M. Knudsen & H. D. Hansen

  2. Faculty of Health and Medical Sciences, University of Copenhagen, 2200, Copenhagen N, Denmark

    L. L. Donovan & G. M. Knudsen

  3. PET and Cyclotron Unit, Copenhagen University Hospital Rigshospitalet, 2100, Copenhagen O, Denmark

    S. Lehel

  4. MGH/HST A. A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Charlestown, MA, 02129, USA

    J. M. Hooker

Authors
  1. L. L. Donovan
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  2. J. H. Magnussen
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  3. A. Dyssegaard
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  4. S. Lehel
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  5. J. M. Hooker
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  6. G. M. Knudsen
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  7. H. D. Hansen
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Corresponding author

Correspondence to H. D. Hansen.

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Conflict of Interest

LLD, GMK, and HDH report grants from The Lundbeck Foundation. JHM, AD, SL declare no conflict of interest. JMH reports grants from NIH, Brain and Behavior Foundation, CureAlz Foundation; non-financial support and other from MGH; personal fees from Rodin Therapeutics, Psy Therapeutics, Merck, NIMH, Evelop Biosciences, Treventis, American Chemical Society, SV Life Sciences, Sunovian, Vertex, Therapeutics; grants and personal fees from Alzheimer’s Drug Discovery Foundation; and other from Eikonizo. In addition, Dr. Hooker has a patent PCT/US2O14/061179 with royalties paid to NucMedCor and Rodin Therapeutics (Previously).

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Donovan, L.L., Magnussen, J.H., Dyssegaard, A. et al. Imaging HDACs In Vivo: Cross-Validation of the [11C]Martinostat Radioligand in the Pig Brain. Mol Imaging Biol 22, 569–577 (2020). https://doi.org/10.1007/s11307-019-01403-9

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  • DOI: https://doi.org/10.1007/s11307-019-01403-9

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