Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats

Vu Anh Truong; Ya Hui Lin; Nuong Thi Kieu Nguyen; Mu Nung Hsu; Nam Ngoc Pham; Yi Hao Chang; Chin Wei Chang; Chih Che Shen; Hsiang Sheng Lee; Po Liang Lai; Yelena V. Parfyonova; Mikhail Menshikov; Jaw Ching Wu; Yu Han Chang; Yu Chen Hu

doi:10.1016/j.ymthe.2021.08.024

Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats

Vu Anh Truong
, Ya Hui Lin
, Nuong Thi Kieu Nguyen
, Mu Nung Hsu
, Nam Ngoc Pham
, Yi Hao Chang
, Chin Wei Chang
, Chih Che Shen
, Hsiang Sheng Lee
, Po Liang Lai
, Yelena V. Parfyonova
, Mikhail Menshikov
, Jaw Ching Wu
, Yu Han Chang^*
, Yu Chen Hu^*

^*Corresponding author for this work

School of Medicine

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

14 Scopus citations

Abstract

Calvarial bone healing is challenging, especially for individuals with osteoporosis because stem cells from osteoporotic patients are highly prone to adipogenic differentiation. Based on previous findings that chondrogenic induction of adipose-derived stem cells (ASCs) can augment calvarial bone healing, we hypothesized that activating chondroinductive Sox Trio genes (Sox5, Sox6, Sox9) and repressing adipoinductive genes (C/ebp-α, Ppar-γ) in osteoporotic ASCs can reprogram cell differentiation and improve calvarial bone healing after implantation. However, simultaneous gene activation and repression in ASCs is difficult. To tackle this problem, we built a CRISPR-BiD system for bi-directional gene regulation. Specifically, we built a CRISPR-AceTran system that exploited both histone acetylation and transcription activation for synergistic Sox Trio activation. We also developed a CRISPR interference (CRISPRi) system that exploited DNA methylation for repression of adipoinductive genes. We combined CRISPR-AceTran and CRISPRi to form the CRISPR-BiD system, which harnessed three mechanisms (transcription activation, histone acetylation, and DNA methylation). After delivery into osteoporotic rat ASCs, CRISPR-BiD significantly enhanced chondrogenesis and in vitro cartilage formation. Implantation of the engineered osteoporotic ASCs into critical-sized calvarial bone defects significantly improved bone healing in osteoporotic rats. These results implicated the potential of the CRISPR-BiD system for bi-directional regulation of cell fate and regenerative medicine.

Original language	English
Pages (from-to)	92-104
Number of pages	13
Journal	Molecular Therapy
Volume	30
Issue number	1
DOIs	https://doi.org/10.1016/j.ymthe.2021.08.024
State	Published - 05 01 2022

Bibliographical note

Publisher Copyright:
© 2021 The American Society of Gene and Cell Therapy

Keywords

CRISPR-AceTran
CRISPR-BiD
CRISPRi
bi-direction gene regulation
regenerative medicine

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10.1016/j.ymthe.2021.08.024

Cite this

Truong, V. A., Lin, Y. H., Nguyen, N. T. K., Hsu, M. N., Pham, N. N., Chang, Y. H., Chang, C. W., Shen, C. C., Lee, H. S., Lai, P. L., Parfyonova, Y. V., Menshikov, M., Wu, J. C., Chang, Y. H., & Hu, Y. C. (2022). Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats. Molecular Therapy, 30(1), 92-104. https://doi.org/10.1016/j.ymthe.2021.08.024

@article{d88d593410b54aeaae9b3ebd4176b756,

title = "Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats",

abstract = "Calvarial bone healing is challenging, especially for individuals with osteoporosis because stem cells from osteoporotic patients are highly prone to adipogenic differentiation. Based on previous findings that chondrogenic induction of adipose-derived stem cells (ASCs) can augment calvarial bone healing, we hypothesized that activating chondroinductive Sox Trio genes (Sox5, Sox6, Sox9) and repressing adipoinductive genes (C/ebp-α, Ppar-γ) in osteoporotic ASCs can reprogram cell differentiation and improve calvarial bone healing after implantation. However, simultaneous gene activation and repression in ASCs is difficult. To tackle this problem, we built a CRISPR-BiD system for bi-directional gene regulation. Specifically, we built a CRISPR-AceTran system that exploited both histone acetylation and transcription activation for synergistic Sox Trio activation. We also developed a CRISPR interference (CRISPRi) system that exploited DNA methylation for repression of adipoinductive genes. We combined CRISPR-AceTran and CRISPRi to form the CRISPR-BiD system, which harnessed three mechanisms (transcription activation, histone acetylation, and DNA methylation). After delivery into osteoporotic rat ASCs, CRISPR-BiD significantly enhanced chondrogenesis and in vitro cartilage formation. Implantation of the engineered osteoporotic ASCs into critical-sized calvarial bone defects significantly improved bone healing in osteoporotic rats. These results implicated the potential of the CRISPR-BiD system for bi-directional regulation of cell fate and regenerative medicine.",

keywords = "CRISPR-AceTran, CRISPR-BiD, CRISPRi, bi-direction gene regulation, regenerative medicine",

author = "Truong, \{Vu Anh\} and Lin, \{Ya Hui\} and Nguyen, \{Nuong Thi Kieu\} and Hsu, \{Mu Nung\} and Pham, \{Nam Ngoc\} and Chang, \{Yi Hao\} and Chang, \{Chin Wei\} and Shen, \{Chih Che\} and Lee, \{Hsiang Sheng\} and Lai, \{Po Liang\} and Parfyonova, \{Yelena V.\} and Mikhail Menshikov and Wu, \{Jaw Ching\} and Chang, \{Yu Han\} and Hu, \{Yu Chen\}",

note = "Publisher Copyright: {\textcopyright} 2021 The American Society of Gene and Cell Therapy",

year = "2022",

month = jan,

day = "5",

doi = "10.1016/j.ymthe.2021.08.024",

language = "英语",

volume = "30",

pages = "92--104",

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Truong, VA, Lin, YH, Nguyen, NTK, Hsu, MN, Pham, NN, Chang, YH, Chang, CW, Shen, CC, Lee, HS, Lai, PL, Parfyonova, YV, Menshikov, M, Wu, JC, Chang, YH & Hu, YC 2022, 'Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats', Molecular Therapy, vol. 30, no. 1, pp. 92-104. https://doi.org/10.1016/j.ymthe.2021.08.024

TY - JOUR

T1 - Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats

AU - Truong, Vu Anh

AU - Lin, Ya Hui

AU - Nguyen, Nuong Thi Kieu

AU - Hsu, Mu Nung

AU - Pham, Nam Ngoc

AU - Chang, Yi Hao

AU - Chang, Chin Wei

AU - Shen, Chih Che

AU - Lee, Hsiang Sheng

AU - Lai, Po Liang

AU - Parfyonova, Yelena V.

AU - Menshikov, Mikhail

AU - Wu, Jaw Ching

AU - Chang, Yu Han

AU - Hu, Yu Chen

PY - 2022/1/5

Y1 - 2022/1/5

N2 - Calvarial bone healing is challenging, especially for individuals with osteoporosis because stem cells from osteoporotic patients are highly prone to adipogenic differentiation. Based on previous findings that chondrogenic induction of adipose-derived stem cells (ASCs) can augment calvarial bone healing, we hypothesized that activating chondroinductive Sox Trio genes (Sox5, Sox6, Sox9) and repressing adipoinductive genes (C/ebp-α, Ppar-γ) in osteoporotic ASCs can reprogram cell differentiation and improve calvarial bone healing after implantation. However, simultaneous gene activation and repression in ASCs is difficult. To tackle this problem, we built a CRISPR-BiD system for bi-directional gene regulation. Specifically, we built a CRISPR-AceTran system that exploited both histone acetylation and transcription activation for synergistic Sox Trio activation. We also developed a CRISPR interference (CRISPRi) system that exploited DNA methylation for repression of adipoinductive genes. We combined CRISPR-AceTran and CRISPRi to form the CRISPR-BiD system, which harnessed three mechanisms (transcription activation, histone acetylation, and DNA methylation). After delivery into osteoporotic rat ASCs, CRISPR-BiD significantly enhanced chondrogenesis and in vitro cartilage formation. Implantation of the engineered osteoporotic ASCs into critical-sized calvarial bone defects significantly improved bone healing in osteoporotic rats. These results implicated the potential of the CRISPR-BiD system for bi-directional regulation of cell fate and regenerative medicine.

AB - Calvarial bone healing is challenging, especially for individuals with osteoporosis because stem cells from osteoporotic patients are highly prone to adipogenic differentiation. Based on previous findings that chondrogenic induction of adipose-derived stem cells (ASCs) can augment calvarial bone healing, we hypothesized that activating chondroinductive Sox Trio genes (Sox5, Sox6, Sox9) and repressing adipoinductive genes (C/ebp-α, Ppar-γ) in osteoporotic ASCs can reprogram cell differentiation and improve calvarial bone healing after implantation. However, simultaneous gene activation and repression in ASCs is difficult. To tackle this problem, we built a CRISPR-BiD system for bi-directional gene regulation. Specifically, we built a CRISPR-AceTran system that exploited both histone acetylation and transcription activation for synergistic Sox Trio activation. We also developed a CRISPR interference (CRISPRi) system that exploited DNA methylation for repression of adipoinductive genes. We combined CRISPR-AceTran and CRISPRi to form the CRISPR-BiD system, which harnessed three mechanisms (transcription activation, histone acetylation, and DNA methylation). After delivery into osteoporotic rat ASCs, CRISPR-BiD significantly enhanced chondrogenesis and in vitro cartilage formation. Implantation of the engineered osteoporotic ASCs into critical-sized calvarial bone defects significantly improved bone healing in osteoporotic rats. These results implicated the potential of the CRISPR-BiD system for bi-directional regulation of cell fate and regenerative medicine.

KW - CRISPR-AceTran

KW - CRISPR-BiD

KW - CRISPRi

KW - bi-direction gene regulation

KW - regenerative medicine

UR - https://www.scopus.com/pages/publications/85114286135

U2 - 10.1016/j.ymthe.2021.08.024

DO - 10.1016/j.ymthe.2021.08.024

M3 - 文章

C2 - 34450254

AN - SCOPUS:85114286135

SN - 1525-0016

VL - 30

SP - 92

EP - 104

JO - Molecular Therapy

JF - Molecular Therapy

IS - 1

ER -

Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats

Abstract

Bibliographical note

Keywords

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