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An Hsp70 promoter–based mouse for heat shock–induced gene modulation

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Abstract

Physical therapy is extensively employed in clinical settings. Nevertheless, the absence of suitable animal models has resulted in an incomplete understanding of the in vivo mechanisms and cellular distribution that respond to physical stimuli. The objective of this research was to create a mouse model capable of indicating the cells affected by physical stimuli. In this study, we successfully established a mouse line based on the heat shock protein 70 (Hsp70) promoter, wherein the expression of CreERT2 can be induced by physical stimuli. Following stimulation of the mouse tail, ear, or cultured calvarias with heat shock (generated by heating, ultrasound, or laser), a distinct Cre-mediated excision was observed in cells stimulated by these physical factors with minimal occurrence of leaky reporter expression. The application of heat shock to Hsp70-CreERT2; FGFR2-P253R double transgenic mice or Hsp70-CreERT2 mice infected with AAV-BMP4 at calvarias induced the activation of Cre-dependent mutant FGFR2-P253R or BMP4 respectively, thereby facilitating the premature closure of cranial sutures or the repair of calvarial defects. This novel mouse line holds significant potential for investigating the underlying mechanisms of physical therapy, tissue repair and regeneration, lineage tracing, and targeted modulation of gene expression of cells in local tissue stimulated by physical factor at the interested time points.

Key messages

  • In the study, an Hsp70-CreERT2 transgenic mouse was generated for heat shock-induced gene modulation.

  • Heat shock, ultrasound, and laser stimulation effectively activated Cre expression in Hsp70-CreERT2; reporter mice, which leads to deletion of floxed DNA sequence in the tail, ear, and cultured calvaria tissues of mice.

  • Local laser stimuli on cultured calvarias effectively induce Fgfr2-P253R expression in Hsp70-mTmG-Fgfr2-P253R mice and result in accelerated premature closure of cranial suture.

  • Heat shock activated AAV9-FLEX-BMP4 expression and subsequently promoted the repair of calvarial defect of Hsp70-CreERT2; Rosa26-mTmG mice.

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Data availability

All data generated or analyzed during this study are included either in this article or in the additional files.

Abbreviations

LIPUS:

Low-intensity pulsed ultrasound

AAV:

Adeno-associated virus

FGFR2:

Fibroblast growth factor receptor type 2

RUNX2:

Runt-related transcription factor 2

BMP4:

Bone morphogenetic protein

OC:

Osteocalcin

OSX:

Osterix

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Funding

National Key Research and Development Program of China, 2018YFA0800802, Lin Chen, National Natural Science Foundation of China, 92168121, Fengtao Luo,82161160313, Lin Chen, 82122044, Yangli Xie, Chongqing Natural Science Foundation, cstc2020jcyj-msxmX0112, Fengtao Luo, CSTB2022NSCQ-MSX0936, Min Jin.

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Author notes

  1. Hangang Chen and Yangli Xie contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Wound Repair and Rehabilitation Medicine, Daping Hospital, Third Military Medical University, Chongqing, 400042, China

    Hangang Chen, Yangli Xie, Mei Zhang, Junlan Huang, Ruobin Zhang, Can Li, Xiaolan Du, Hua Chen, Qiang Nie, Sen Liang, Qiaoyan Tan, Jing Yang, Min Jin, Shuo Huang, Liang Kuang, Nan Su, Huabing Qi, Xiaoqing Luo, Lin Chen & Fengtao Luo

  2. State Key Laboratory of Trauma, Burn and Combined Injury, Third Military Medical University, Chongqing, 400038, China

    Yangli Xie, Xiaolan Du, Qiaoyan Tan, Jing Yang, Min Jin, Nan Su, Huabing Qi, Lin Chen & Fengtao Luo

  3. Department of Chinese Medicine Rehabilitation, Chongqing Emergency Medical Center, Chongqing University Central Hospital), Chongqing, 400042, China

    Wanling Jiang

  4. Faculty of Health Sciences, University of Macau, Macau SAR, China

    Xiaoling Xu & Chuxia Deng

Authors
  1. Hangang Chen
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Contributions

Study design: F.L., Y.X., and L.C.; study conduct: H.C., M.Z., J.H., R.Z., H.C., Q.N., C.L., N.S., S.L., and W.J; data collection: M.J. K.L., J.Y., Q.T., X.L., and S.H.; data analysis and interpretation: F.L, Y.X., H.Q., X.X., C.D., and X.D.; drafting the manuscript: H.C., F. L, Y.X., and L.C. All the authors read and approved the final paper.

Corresponding authors

Correspondence to Lin Chen or Fengtao Luo.

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Animal experiments were approved by the Institutional Animal Care and Use Committee of Daping Hospital (Chongqing, China).

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Chen, H., Xie, Y., Zhang, M. et al. An Hsp70 promoter–based mouse for heat shock–induced gene modulation. J Mol Med 102, 693–707 (2024). https://doi.org/10.1007/s00109-024-02433-9

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