Background This study investigated the role and underlying mechanism of large tumor suppressor 1/2 (LATS1/2), a core kinase in the Hippo pathway, in inhibiting osteoclastic differentiation and ameliorating ovariectomy (OVX)-induced osteoporosis in rats through the RANKL-dependent Hippo–YAP/TAZ signaling pathway both in vivo and in vitro.
Methods Under stimulation by receptor activator of nuclear factor-κB ligand (RANKL), small interfering RNA (siRNA) was used to suppress the expression of LATS1/2 and YAP. The levels of osteoclast-related genes and proteins, as well as YAP pathway proteins, were evaluated in RAW264.7 cells and bone marrow-derived macrophages (BMMs) via quantitative reverse transcription polymerase chain reaction and Western blot analysis. Cell viability and osteoclast formation potential were measured via the Cell Counting Kit-8 assay and tartrate-resistant acid phosphatase (TRAP) staining, respectively. Osteoclasts and filamentous actin were stained with TRAP and phalloidin to characterize cell morphology. Furthermore, to research the role of the Hippo/YAP pathway, we used siRNA to knock down LATS1/2 and examined its effect on RANKL-mediated osteoclast differentiation. In addition, we constructed an OVX-induced osteoporosis model in 8-week-old female Sprague‒Dawley rats, treated them with a LATS1/2 inhibitor for 4 weeks at 12 weeks after surgery, and evaluated the remission of osteoporosis by micro-CT and histological methods with a LATS1/2 inhibitor.
Results After LATS1/2 was silenced or inhibited in BMMs and RAW264.7 cells, YAP was significantly upregulated and YAP phosphorylation and osteoclast function were inhibited. By interfering with YAP expression, the Hippo/YAP pathway inhibited osteoclast differentiation in BMMs. In vivo, injection of LATS1/2 inhibitor alleviated osteoporosis in ovariectomized rats.
Conclusion Our findings indicate that the inhibition of LATS1/2 considerably suppresses osteoclast differentiation and alleviates OVX-induced osteoporosis in rats. Additionally, our results show that LATS1/2 inhibitors are important for the differentiation of osteoclasts and highlight the possible therapeutic value of LATS1/2 inhibitors.
| [1] |
Salhotra A, Shah HN, Levi B, et al. . Mechanisms of bone development and repair. Nat Rev Mol Cell Biol., 2020, 21(11): 696-711
|
| [2] |
Zhao X, Patil S, Xu F, et al. . Role of biomolecules in osteoclasts and their therapeutic potential for osteoporosis. Biomolecules., 2021, 11(5): 747
|
| [3] |
Gao Y, Patil S, Jia J. The development of molecular biology of osteoporosis. Int J Mol Sci., 2021, 22(15): 8182
|
| [4] |
Cao X. RANKL-RANK signaling regulates osteoblast differentiation and bone formation. Bone Res., 2018, 6: 35
|
| [5] |
Li Y, Zhuang Q, Tao L, et al. . Urolithin B suppressed osteoclast activation and reduced bone loss of osteoporosis via inhibiting ERK/NF-κB pathway. Cell Prolif., 2022, 55(10): e13291
|
| [6] |
Chen K, Qiu P, Yuan Y, et al. . Pseurotin a inhibits osteoclastogenesis and prevents ovariectomized-induced bone loss by suppressing reactive oxygen species. Theranostics., 2019, 9(6): 1634-1650
|
| [7] |
Abe Y, Kofman ER, Almeida M, et al. . RANK ligand converts the NCoR/HDAC3 co-repressor to a PGC1β- and RNA-dependent co-activator of osteoclast gene expression. Mol Cell., 2023, 83(19): 3421-3437.e11
|
| [8] |
Meng Z, Li FL, Fang C, et al. . The Hippo pathway mediates Semaphorin signaling. Sci Adv., 2022, 8(21): eabl9806
|
| [9] |
Yu FX, Zhao B, Guan KL. Hippo pathway in organ size control, tissue homeostasis, and cancer. Cell., 2015, 163(4): 811-828
|
| [10] |
Zheng Y, Pan D. The hippo signaling pathway in development and disease. Dev Cell., 2019, 50(3): 264-282
|
| [11] |
Moroishi T, Hayashi T, Pan WW, et al. . The hippo pathway kinases LATS1/2 suppress cancer immunity. Cell., 2016, 167(6): 1525-1539.e17
|
| [12] |
Sun K, Guo J, Guo Z, et al. . The roles of the Hippo-YAP signalling pathway in Cartilage and Osteoarthritis. Ageing Res Rev., 2023, 90: 102015
|
| [13] |
Lorthongpanich C, Thumanu K, Tangkiettrakul K, et al. . YAP as a key regulator of adipo-osteogenic differentiation in human MSCs. Stem Cell Res Ther., 2019, 10(1): 402
|
| [14] |
Park JS, Kim M, Song NJ, et al. . A reciprocal role of the Smad4-taz axis in osteogenesis and adipogenesis of mesenchymal stem cells. Stem Cells., 2019, 37(3): 368-381
|
| [15] |
Li L, Dong L, Wang Y, et al. . Lats1/2-mediated alteration of hippo signaling pathway regulates the fate of bone marrow-derived mesenchymal stem cells. Biomed Res Int., 2018, 2018: 4387932
|
| [16] |
Black DM, Rosen CJ. Postmenopausal osteoporosis. N Engl J Med., 2016, 374(3): 254-262
|
| [17] |
Yu B, Wang CY. Osteoporosis and periodontal diseases–An update on their association and mechanistic links. Periodontol 2000., 2022, 89(1): 99-113
|
| [18] |
Novack DV, Teitelbaum SL. The osteoclast: friend or foe?. Annu Rev Pathol Mech Dis., 2008, 3: 457-484
|
| [19] |
Boyce BF. Advances in the regulation of osteoclasts and osteoclast functions. J Dent Res., 2013, 92(10): 860-867
|
| [20] |
Takegahara N, Kim H, Choi Y. Unraveling the intricacies of osteoclast differentiation and maturation: insight into novel therapeutic strategies for bone-destructive diseases. Exp Mol Med., 2024, 56(2): 264-272
|
| [21] |
Yang W, Han W, Qin A, et al. . The emerging role of Hippo signaling pathway in regulating osteoclast formation. J Cell Physiol., 2018, 233(6): 4606-4617
|
| [22] |
Hong AW, Meng Z, Guan KL. The Hippo pathway in intestinal regeneration and disease. Nat Rev Gastroenterol Hepatol., 2016, 13(6): 324-337
|
| [23] |
Zanconato F, Cordenonsi M, Piccolo S. YAP/TAZ at the roots of cancer. Cancer Cell., 2016, 29(6): 783-803
|
| [24] |
Moya IM, Halder G. Hippo–YAP/TAZ signalling in organ regeneration and regenerative medicine. Nat Rev Mol Cell Biol., 2019, 20(4): 211-226
|
| [25] |
Xiong J, Almeida M, O’Brien CA. The YAP/TAZ transcriptional co-activators have opposing effects at different stages of osteoblast differentiation. Bone., 2018, 112: 1-9
|
| [26] |
Tang Y, Feinberg T, Keller ET, et al. . Snail/Slug binding interactions with YAP/TAZ control skeletal stem cell self-renewal and differentiation. Nat Cell Biol., 2016, 18(9): 917-929
|
| [27] |
Yang W, Lu X, Zhang T, et al. . TAZ inhibits osteoclastogenesis by attenuating TAK1/NF-κB signaling. Bone Res., 2021, 9: 33
|
| [28] |
Zheng C, Liu H, Zhao P, et al. . Targeting sulfation-dependent mechanoreciprocity between matrix and osteoblasts to mitigate bone loss. Sci Transl Med., 2023, 15(710): eadg3983
|
| [29] |
Kegelman CD, Mason DE, Dawahare JH, et al. . Skeletal cell YAP and TAZ combinatorially promote bone development. FASEB J., 2018, 32(5): 2706-2721
|
| [30] |
Kegelman CD, Collins JM, Nijsure MP, et al. . Gone caving: roles of the transcriptional regulators YAP and TAZ in skeletal development. Curr Osteoporos Rep., 2020, 18(5): 526-540
|
| [31] |
Meng Z, Moroishi T, Mottier-Pavie V, et al. . MAP4K family kinases act in parallel to MST1/2 to activate LATS1/2 in the Hippo pathway. Nat Commun., 2015, 6: 8357
|
| [32] |
Kern JG, Tilston-Lunel AM, Federico A, et al. . Inactivation of LATS1/2 drives luminal-basal plasticity to initiate basal-like mammary carcinomas. Nat Commun., 2022, 13(1): 7198
|
| [33] |
Pan WW, Moroishi T, Koo JH, et al. . Cell type-dependent function of LATS1/2 in cancer cell growth. Oncogene., 2019, 38(14): 2595-2610
|
| [34] |
Furth N, Aylon Y. The LATS1 and LATS2 tumor suppressors: beyond the Hippo pathway. Cell Death Differ., 2017, 24(9): 1488-1501
|
| [35] |
Qi S, Zhu Y, Liu X, et al. . WWC proteins mediate LATS1/2 activation by Hippo kinases and imply a tumor suppression strategy. Mol Cell., 2022, 82(10): 1850-1864.e7
|
| [36] |
Yang Q, Lv Z, Wang M, et al. . LATS1/2 loss promote tumor immune evasion in endometrial cancer through downregulating MHC-I expression. J Exp Clin Cancer Res., 2024, 43(1): 54
|
| [37] |
Li K, Liu L, Liu H, et al. . LATS1/YAP1 axis controls bone regeneration on distraction osteogenesis by activating Wnt/β-catenin. Tissue Eng Part A., 2024, 30(3–4): 154-167
|
| [38] |
Li K, Liu L, Liu H, et al. . Hippo/YAP1 promotes osteoporotic mice bone defect repair via the activating of Wnt signaling pathway. Cell Signal., 2024, 116: 111037
|
| [39] |
Rothzerg E, Ingley E, Mullin B, Xue W, Wood D, Xu J. The Hippo in the room: Targeting the Hippo signaling pathway for osteosarcoma therapies. J Cell Physiol., 2021, 236(3): 1606-1615
|
Funding
the First Hospital of Jiaxing(2022-xkdtr-01)
the First Hospital of Jiaxing (2024-QMX-013)
Science and Technology Bureau of Jiaxing City(2022-yzcsgtjzjz)
RIGHTS & PERMISSIONS
The Author(s), under exclusive licence to the Huazhong University of Science and Technology
PDF
