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1. Mechanical Stretch Control of Adipocyte AKT Signaling and the Role of FAK and ROCK Mechanosensors.

Author

Bouzid, Tasneem, Kim, Eunju, Riehl, Brandon D., Yang, Ruiguo, Saraswathi, Viswanathan, Kim, Jason K., and Lim, Jung Yul

Subjects
FOCAL adhesion kinase, CYCLIC loads, ADIPOSE tissues, FOCAL adhesions, TYPE 2 diabetes, INSULIN
Abstract

Adipose tissue in vivo is physiologically exposed to compound mechanical loading due to bodyweight bearing, posture, and motion. The capability of adipocytes to sense and respond to mechanical loading milieus to influence metabolic functions may provide a new insight into obesity and metabolic diseases such as type 2 diabetes (T2D). Here, we evidenced physiological mechanical loading control of adipocyte insulin signaling cascades. We exposed differentiated 3T3-L1 adipocytes to mechanical stretching and assessed key markers of insulin signaling, AKT activation, and GLUT4 translocation, required for glucose uptake. We showed that cyclic stretch loading at 5% strain and 1 Hz frequency increases AKT phosphorylation and GLUT4 translocation to the plasma membrane by approximately two-fold increases compared to unstretched controls for both markers as assessed by immunoblotting (p < 0.05). These results indicate that cyclic stretching activates insulin signaling and GLUT4 trafficking in adipocytes. In the mechanosensing mechanism study, focal adhesion kinase (FAK) inhibitor (FAK14) and RhoA kinase (ROCK) inhibitor (Y-27632) impaired actin cytoskeleton structural formation and significantly suppressed the stretch induction of AKT phosphorylation in adipocytes (p < 0.001). This suggests the regulatory role of focal adhesion and cytoskeletal mechanosensing in adipocyte insulin signaling under stretch loading. Our finding on the impact of mechanical stretch loading on key insulin signaling effectors in differentiated adipocytes and the mediatory role of focal adhesion and cytoskeleton mechanosensors is the first of its kind to our knowledge. This may suggest a therapeutic potential of mechanical loading cue in improving conditions of obesity and T2D. For instance, cyclic mechanical stretch loading of adipose tissue could be explored as a tool to improve insulin sensitivity in patients with obesity and T2D, and the mediatory mechanosensors such as FAK and ROCK may be targeted to further invigorate stretch-induced insulin signaling activation. [ABSTRACT FROM AUTHOR]

Published
2024
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7. List of Contributors

Author

Ahmed, Abu S.I., primary, Anversa, Piero, additional, Apoorva, Fnu, additional, Arakawa, Christopher K., additional, Baylink, David J., additional, Benameur, Laila, additional, Bernhard, Jonathan, additional, Bhatia, Mickie, additional, Blatchley, Michael, additional, Borenstein, Jeffrey T., additional, Brandenberg, Nathalie, additional, Breault, David T., additional, Brun, Caroline E., additional, Carrier, Rebecca L., additional, Cetinbas, Naniye Malli, additional, Chen, Wanqiu, additional, Cheng, Yu-Hao, additional, Chevalier, Fabien P., additional, Clevers, Hans, additional, Conboy, Michael J., additional, Conboy, Irina M., additional, Conover, Joanne C., additional, DeForest, Cole A., additional, Donahue, Henry J., additional, Dumont, Nicolas A., additional, Ferlin, Kimberly M., additional, Findlay, Michael W., additional, Fisher, John P., additional, Freudenberg, Uwe, additional, Funaki, Makoto, additional, Gerecht, Sharon, additional, Goichberg, Polina, additional, Griffith, Linda G., additional, Guasch, Géraldine, additional, Guild, Joshua, additional, Guo, Ting, additional, Gurtner, Geoffrey C., additional, Habibovic, Pamela, additional, Haque, Amranul, additional, He, Xi C., additional, Hernandez-Gordillo, Victor, additional, Hosoda, Toru, additional, Huang, Jie, additional, Ito, Yoshihiro, additional, Janmey, Paul A., additional, Jiang, Lei, additional, Jones, Peter Anthony, additional, Kaplan, David S., additional, Karp, Jeffrey M., additional, Klecker, Christina, additional, Koppes, Abigail N., additional, Krause, Arthur, additional, Leena, Maria, additional, Leri, Annarosa, additional, Levenberg, Shulamit, additional, Li, Xiaowei, additional, Li, Yingying, additional, Li, Yan, additional, Li, Linheng, additional, Lim, Jung Yul, additional, Liu, Yijun, additional, Lutolf, Matthias P., additional, Ma, Teng, additional, Maeda, Kay, additional, Malhotra, Angad, additional, Manivasagam, Geetha, additional, Mao, Hongli, additional, Mao, Hai-Quan, additional, McDevitt, Todd C., additional, Mekhail, Mina, additional, Moccetti, Tiziano, additional, Müller, Eike, additional, Nair, Lakshmi S., additional, Nakanishi, Mio, additional, Ng, Johnathan, additional, Pasricha, Renu, additional, Perry, John, additional, Pompe, Tilo, additional, Ramalingam, Murugan, additional, Ramasamy, Keerthana, additional, Rana, Deepti, additional, Revzin, Alexander, additional, Riehl, Brandon D., additional, Roman, Jose, additional, Roper, Jatin, additional, Rosenfeld, Dekel, additional, Rota, Marcello, additional, Rouwkema, Jeroen, additional, Rudnicki, Michael A., additional, Ruel, Marc, additional, Saez, Borja, additional, Sasaki, Nobuo, additional, Sato, Toshiro, additional, Scadden, David T., additional, Shroff, Sanaya N., additional, Singh, Ankur, additional, Smith, Quinton, additional, Spiller, Kara, additional, Suuronen, Erik J., additional, Tabrizian, Maryam, additional, Tang, Xiaolei, additional, Todd, Krysti L., additional, Tsai, Ang-Chen, additional, van Blitterswijk, Clemens, additional, Venkatraman, Aparna, additional, Vishwakarma, Ajaykumar, additional, Vunjak-Novakovic, Gordana, additional, Wang, Jane, additional, Wang, Shutao, additional, Wasnik, Samiksha, additional, Werner, Carsten, additional, Wilson, Jenna L., additional, Yılmaz, Ömer H., additional, Yuan, Xuegang, additional, Yusuf, Rushdia Z., additional, Zhang, Xiao-Bing, additional, and Zhao, Meng, additional

Published
2017
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9. List of Contributors

Author

Ahadian, Samad, primary, Akiyaman, Kentaro, additional, Alansari, Sarah, additional, Alikhani, Mani, additional, Alikhani, Mona, additional, Arthur, Agnieszka, additional, Arulkumar, Shylaja, additional, Baiguera, Silvia, additional, Bianco, Alessandra, additional, Bissada, Nabil F., additional, Bleicher, Françoise, additional, Bliley, Jacqueline M., additional, Bosshardt, Dieter D., additional, Botero, Tatiana M., additional, Braga, Raquel, additional, Briggs, Kristen K., additional, Brooks, Patricia J., additional, Cannon, Kevin, additional, Carrouel, Florence, additional, Chavez, Miquella G., additional, Cheng, Ming-Te, additional, Christ, George J., additional, Cook, Colin A., additional, Cooper, Paul R., additional, Cox, Timothy C., additional, Cunningham, Michael L., additional, Dashe, Jesse, additional, Davidovitch, Ze’ev, additional, Davies, Lindsay C., additional, Del Gaudio, Costantino, additional, Diekwisch, Thomas G.H., additional, Diogenes, Anibal, additional, Duncan, Randall L., additional, Edwards, Paul C., additional, El-Bialy, Tarek, additional, Enlow, Donald H., additional, Farach-Carson, Mary C., additional, Farges, Jean-Christophe, additional, Feinberg, Stephen E., additional, Felfy, Hady, additional, Freeman, Phillip N., additional, Freilich, Martin, additional, Fujie, Toshinori, additional, Gao, Changyou, additional, Giannobile, William V., additional, Gong, Siew-Ging, additional, Grayson, Warren L., additional, Greene, Robert M., additional, Gronthos, Stan, additional, Gruber, Reinhard, additional, Häkkinen, Lari, additional, Hanna, Craig, additional, Hargreaves, Kenneth M., additional, Satoshi, Emilio Hara, additional, Harrington, Daniel A., additional, Hashmi, Basma, additional, Heino, Jyrki, additional, Hing, Anne V., additional, Holmes, Christina, additional, Honda, Masaki J., additional, Horst, Jeremy A., additional, Hu, Michael S., additional, Hughes, Francis J., additional, Hung, Ben P., additional, Huri, Pinar Yilgor, additional, Ingber, Donald E., additional, Isokawa, Keitaro, additional, Izumi, Kenji, additional, James, Eric N., additional, Jia, Xinqiao, additional, Jin, Yan, additional, Emily Joo, E., additional, Juuri, Emma, additional, Kagami, Hideaki, additional, Kaji, Hirokazu, additional, Kang, Mo K., additional, Kashi, A., additional, Kato, Hiroko, additional, Kaur, Jasvir, additional, Keller, Letitia V., additional, Kemp, Paul D., additional, Khademhosseini, Ali, additional, Khoo, Edmund, additional, Kim, Hae-Won, additional, King, William J., additional, Kirschner, Richard E., additional, Klein, Ophir D., additional, Knowles, Jonathan C., additional, Koivisto, Leeni, additional, Kökten, Tunay, additional, Krebsbach, Paul H., additional, Kuboki, Takuo, additional, Kuchler-Bopp, Sabine, additional, Kucska, Simone, additional, Kuhn, Liisa, additional, Larjava, Hannu, additional, Lee, Eun-Jung, additional, Lee, Joo-Hee, additional, Lee, Oscar K., additional, Lee, Yoo Bin, additional, Lesot, Hervé, additional, Lewis, Mark P., additional, Li, Bei, additional, Li, Luyan, additional, Lim, Jung Yul, additional, Liu, Xing, additional, Liu, Xiaohua, additional, Lo, David D., additional, Longaker, Michael T., additional, Peter Lorenz, H., additional, Lott, David G., additional, Lungová, Vlasta, additional, Ma, Lie, additional, Macchiarini, Paolo, additional, Mammoto, Tadanori, additional, Marei, Mona K., additional, Marolt, Darja, additional, Marra, Kacey G., additional, Martin, Neil R.W., additional, Matalová, Eva, additional, Matsue, Tomokazu, additional, Mehrazarin, Shebli, additional, Mina, Mina, additional, Modo, Michel, additional, Mokry, Jaroslav, additional, Morsczeck, Christian, additional, Moussa, Rania M., additional, Murphy, Neal C., additional, Mushegyan, Vagan, additional, Nagy, Naglaa B., additional, Nair, Lakshmi S., additional, Nayar, Harry, additional, Nör, Jacques E., additional, Obregón, Raquel, additional, Oh, Tae-Ju, additional, Oka, Kyoko, additional, Ostrovidov, Serge, additional, Park, No-Hee, additional, Passineau, Michael J., additional, Passipieri, Juliana A., additional, Pisal, Rishikaysh, additional, Player, Darren, additional, Pradhan-Bhatt, Swati, additional, Parthiban, Selvakumar Prakash, additional, Prochazka, Jan, additional, Prochazkova, Michaela, additional, Qu, Tiejun, additional, Ramalingam, Murugan, additional, Ramón-Azcón, Javier, additional, Rana, Deepti, additional, Ray, Herbert L., additional, Reed, David A., additional, Reinhardt, Dieter P., additional, Richard, Béatrice, additional, Riehl, Brandon D., additional, Rios, Hector F., additional, Peter Rubin, J., additional, Saad, Manal M., additional, Sachar, Ashneet, additional, Sah, Robert L., additional, Saha, Subrata, additional, Saito, Yoko, additional, Sampathkumar, Kaarunya, additional, Sangsuwon, Chinapa, additional, Seo, Byoung Moo, additional, Sharpe, Paul, additional, Shi, Songtao, additional, Shih, Yu-Ru V., additional, Shiku, Hitoshi, additional, Shin, Seung Yun, additional, Sjöqvist, Sebastian, additional, Sloan, Alastair J., additional, Smith, Anthony (Tony) J., additional, Song, In Seok, additional, Stephens, Neil, additional, Stephens, Phil, additional, Svoboda, Kathy K.H., additional, Tabrizian, Maryam, additional, Teixeira, Cristina, additional, Teixeira, Fabricio B., additional, Temple, Joshua P., additional, Thesleff, Irma, additional, Thivichon-Prince, Béatrice, additional, Toriumi, Taku, additional, Treasure, Trevor E., additional, Um, Soyoun, additional, Vishwakarma, Ajaykumar, additional, Wang, Chunfen, additional, Watson, Deborah, additional, Watson, Jeffrey B., additional, Wen, Bo, additional, Witt, Robert L., additional, Wong, Mark E., additional, Yamada, Kenneth M., additional, Yassin, Hala H., additional, Zakheim, Daniel, additional, Zhang, Bing, additional, and Zimmermann, Andrew S., additional

Published
2015
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18. Macro and Microfluidic Flows for Skeletal Regenerative Medicine.

Author

Riehl, Brandon D. and Jung Yul Lim

Subjects
MICROFLUIDICS, CARTILAGE cells, MECHANOTRANSDUCTION (Cytology), HEMOSTASIS, TISSUE engineering, BONE cells
Abstract

Fluid flow has a great potential as a cell stimulatory tool for skeletal regenerative medicine, because fluid flow-induced bone cell mechanotransduction in vivo plays a critical role in maintaining healthy bone homeostasis. Applications of fluid flow for skeletal regenerative medicine are reviewed at macro and microscale. Macroflow in two dimensions (2D), in which flow velocity varies along the normal direction to the flow, has explored molecular mechanisms of bone forming cell mechanotransduction responsible for flow-regulated differentiation, mineralized matrix deposition, and stem cell osteogenesis. Though 2D flow set-ups are useful for mechanistic studies due to easiness in in situ and post-flow assays, engineering skeletal tissue constructs should involve three dimensional (3D) flows, e.g., flow through porous scaffolds. Skeletal tissue engineering using 3D flows has produced promising outcomes, but 3D flow conditions (e.g., shear stress vs. chemotransport) and scaffold characteristics should further be tailored. Ideally, data gained from 2D flows may be utilized to engineer improved 3D bone tissue constructs. Recent microfluidics approaches suggest a strong potential to mimic in vivo microscale interstitial flows in bone. Though there have been few microfluidics studies on bone cells, it was demonstrated that microfluidic platform can be used to conduct high throughput screening of bone cell mechanotransduction behavior under biomimicking flow condit ions. [ABSTRACT FROM AUTHOR]

Published
2012
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19. Fluid Flow-Induced Mesenchymal Stem Cell Migration: Role of FAK and ROCK Mechanosensors

Author

Riehl, Brandon D

Subjects
Stem cell, Shear, FAK, ROCK, Tissue engineering, Cell therapy, Migration, Mechanotransduction, Regenerative medicine, Molecular, Cellular, and Tissue Engineering, Other Mechanical Engineering
Abstract

The study of mesenchymal stem cell (MSC) migration under mechanical stimulation conditions with investigation of the underlying molecular mechanism could lead to a better understanding and outcomes in stem cell-based regenerative medicine. MSCs having multipotent regenerative capability exist in niches in the bone marrow, muscle, vasculature, and in other tissues throughout the body, and their migration through tissues and vasculature for the repair of damaged tissue is a key process of cell and tissue homeostasis, remodeling, and regeneration. While cell migration in response to cytokines and other chemo-attractants is relatively well understood, little is revealed in regard to the effect of mechanical cues. In this study, we investigated the migration of C3H10T1/2 murine MSCs in response to fluid flow-induced shear stress in vitro. MSCs were subjected to steady flows with physiologically relevant shear stresses of 2, 15, and 25 dyne/cm2 and compared with static control. Fluid shear induced cell migration following the flow direction, which effect was greater at higher shear stresses. To test the molecular mechanism, MSCs with stable knockdown of focal adhesion kinase (FAK) and RhoA kinase (ROCK), each constituting the key component of focal adhesion signaling and cytoskeletal tension signaling respectively, were fluid-sheared. FAK-silenced MSCs showed decreases in fluid shear-induced migration, for example, decreases in migration length, confinement ratio, and motility coefficient. Interestingly, in the presence of ROCK silencing, MSCs were more responsive to fluid shear, showing increases in such migration parameters. Our data may suggest a different role of focal adhesion and cytoskeletal tension in mechanical induction of MSC migration. Adviser: Jung Yul Lim

Published
2014

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