Development of a High-Throughput Lysyl Hydroxylase (LH) Assay and Identification of Small-Molecule Inhibitors against LH2

Ashwini K. Devkota; John R. Veloria; Hou Fu Guo; Jonathan M. Kurie; Eun Jeong Cho; Kevin N. Dalby

doi:10.1177/2472555218817057

Development of a High-Throughput Lysyl Hydroxylase (LH) Assay and Identification of Small-Molecule Inhibitors against LH2

Ashwini K. Devkota, John R. Veloria, Hou Fu Guo, Jonathan M. Kurie, Eun Jeong Cho, Kevin N. Dalby

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

Lysyl hydroxylase-2 (LH2) catalyzes the hydroxylation of telopeptidyl lysine residues on collagen, leading to the formation of stable collagen cross-links that connect collagen molecules and stabilize the extracellular matrix. High levels of LH2 have been reported in the formation and stabilization of hydroxylysine aldehyde-derived collagen cross-links (HLCCs), leading to fibrosis and cancer metastasis in certain tissues. Identification of small-molecule inhibitors targeting LH2 activity requires a robust and suitable assay system, which is currently lacking. Thus, despite being a promising target for these diseases, small-molecule inhibitors for LH2 have yet to be reported. Therefore, we developed a luminescence-based strategy to monitor LH activity and validated its ability to identify new inhibitors in a screen of approximately 65,000 compounds against LH2. Primary hits were confirmed using the same LH assay against mimiviral L230. This newly developed LH assay is robust, suitable for high-throughput screening, and able to identify potent specific inhibitors of LH2.

Original language	English
Pages (from-to)	484-491
Number of pages	8
Journal	SLAS Discovery
Volume	24
Issue number	4
DOIs	https://doi.org/10.1177/2472555218817057
State	Published - Apr 1 2019

Bibliographical note

Publisher Copyright:
© 2018 Society for Laboratory Automation and Screening.

Funding

The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by the Cancer Prevention and Research Institute of Texas (CPRIT), grants RP110532-P1 and RP160657, and by the Welch Foundation, grant F-1390.

Funders	Funder number
National Childhood Cancer Registry – National Cancer Institute	K99CA225633
Welch Foundation	F-1390
Cancer Prevention and Research Institute of Texas	RP160657, RP110532-P1

Keywords

cancer
high-throughput screen
luminescence
lysyl hydroxylase-2 (LH2)
succinate detection

ASJC Scopus subject areas

Biotechnology
Analytical Chemistry
Biochemistry
Molecular Medicine

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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title = "Development of a High-Throughput Lysyl Hydroxylase (LH) Assay and Identification of Small-Molecule Inhibitors against LH2",

abstract = "Lysyl hydroxylase-2 (LH2) catalyzes the hydroxylation of telopeptidyl lysine residues on collagen, leading to the formation of stable collagen cross-links that connect collagen molecules and stabilize the extracellular matrix. High levels of LH2 have been reported in the formation and stabilization of hydroxylysine aldehyde-derived collagen cross-links (HLCCs), leading to fibrosis and cancer metastasis in certain tissues. Identification of small-molecule inhibitors targeting LH2 activity requires a robust and suitable assay system, which is currently lacking. Thus, despite being a promising target for these diseases, small-molecule inhibitors for LH2 have yet to be reported. Therefore, we developed a luminescence-based strategy to monitor LH activity and validated its ability to identify new inhibitors in a screen of approximately 65,000 compounds against LH2. Primary hits were confirmed using the same LH assay against mimiviral L230. This newly developed LH assay is robust, suitable for high-throughput screening, and able to identify potent specific inhibitors of LH2.",

keywords = "cancer, high-throughput screen, luminescence, lysyl hydroxylase-2 (LH2), succinate detection",

author = "Devkota, {Ashwini K.} and Veloria, {John R.} and Guo, {Hou Fu} and Kurie, {Jonathan M.} and Cho, {Eun Jeong} and Dalby, {Kevin N.}",

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doi = "10.1177/2472555218817057",

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AU - Devkota, Ashwini K.

AU - Veloria, John R.

AU - Guo, Hou Fu

AU - Kurie, Jonathan M.

AU - Cho, Eun Jeong

AU - Dalby, Kevin N.

PY - 2019/4/1

Y1 - 2019/4/1

N2 - Lysyl hydroxylase-2 (LH2) catalyzes the hydroxylation of telopeptidyl lysine residues on collagen, leading to the formation of stable collagen cross-links that connect collagen molecules and stabilize the extracellular matrix. High levels of LH2 have been reported in the formation and stabilization of hydroxylysine aldehyde-derived collagen cross-links (HLCCs), leading to fibrosis and cancer metastasis in certain tissues. Identification of small-molecule inhibitors targeting LH2 activity requires a robust and suitable assay system, which is currently lacking. Thus, despite being a promising target for these diseases, small-molecule inhibitors for LH2 have yet to be reported. Therefore, we developed a luminescence-based strategy to monitor LH activity and validated its ability to identify new inhibitors in a screen of approximately 65,000 compounds against LH2. Primary hits were confirmed using the same LH assay against mimiviral L230. This newly developed LH assay is robust, suitable for high-throughput screening, and able to identify potent specific inhibitors of LH2.

AB - Lysyl hydroxylase-2 (LH2) catalyzes the hydroxylation of telopeptidyl lysine residues on collagen, leading to the formation of stable collagen cross-links that connect collagen molecules and stabilize the extracellular matrix. High levels of LH2 have been reported in the formation and stabilization of hydroxylysine aldehyde-derived collagen cross-links (HLCCs), leading to fibrosis and cancer metastasis in certain tissues. Identification of small-molecule inhibitors targeting LH2 activity requires a robust and suitable assay system, which is currently lacking. Thus, despite being a promising target for these diseases, small-molecule inhibitors for LH2 have yet to be reported. Therefore, we developed a luminescence-based strategy to monitor LH activity and validated its ability to identify new inhibitors in a screen of approximately 65,000 compounds against LH2. Primary hits were confirmed using the same LH assay against mimiviral L230. This newly developed LH assay is robust, suitable for high-throughput screening, and able to identify potent specific inhibitors of LH2.

KW - cancer

KW - high-throughput screen

KW - luminescence

KW - lysyl hydroxylase-2 (LH2)

KW - succinate detection

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Development of a High-Throughput Lysyl Hydroxylase (LH) Assay and Identification of Small-Molecule Inhibitors against LH2

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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