Protein kinase B/AKT isoform 2 drives migration of human mesenchymal stem cells

Bulj,Zrinka; Duchi,Serena; Bevilacqua,Alessandro; Gherardi,Alessandro; Dozza,Barbara; Piccinini,Filippo; Adalgisa Mariani,Giulia; Lucarelli,Enrico; Giannini,Sandro; Donati,Davide; Marmiroli,Sandra

doi:10.3892/ijo.2012.1700

Protein kinase B/AKT isoform 2 drives migration of human mesenchymal stem cells

Authors:
- Zrinka Bulj
- Serena Duchi
- Alessandro Bevilacqua
- Alessandro Gherardi
- Barbara Dozza
- Filippo Piccinini
- Giulia Adalgisa Mariani
- Enrico Lucarelli
- Sandro Giannini
- Davide Donati
- Sandra Marmiroli
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Affiliations: Cellular Signalling Laboratory, Department of Biomedical Sciences, 40126 Bologna, Italy, Osteoarticular Regeneration Laboratory, Rizzoli Orthopaedic Institute, 40136 Bologna, Italy, Advanced Research Center on Electronic Systems for Information and Communication Technologies ‘E. De Castro’ (ARCES), 40125 Bologna, Italy, Cellular Signalling Laboratory, Department of Surgery, Medicine, Dentistry and Morphological Sciences, 41124 Modena, Italy
Published online on: November 15, 2012 https://doi.org/10.3892/ijo.2012.1700
Pages: 118-126
Copyright: © Bulj et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

This study was designed to investigate the migratory behavior of adult human mesenchymal stem cells (MSC) and the underlying mechanism. Cell migration was assessed by transwell, wound healing and time-lapse in vivo motility assays. Pharmacological inhibitors were used to determine the potential mechanism responsible for cell migration and invasion. The tests that were implemented revealed that MSC were fairly migratory. Protein kinase B (AKT) was strongly activated at the basal level. Through our analyses we demonstrated that pharmacological inactivation of AKT2 but not AKT1 significantly decreased cell migration and invasion. Although preliminary, collectively our results indicate that AKT2 activation plays a critical role in enabling MSC migration.

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1.	Charbord P: Bone marrow mesenchymal stem cells: historical overview and concepts. Hum Gene Ther. 21:1045–1056. 2010. View Article : Google Scholar : PubMed/NCBI
2.	Devine SM, Cobbs C, Jennings M, Bartholomew A and Hoffman R: Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood. 101:2999–3001. 2010. View Article : Google Scholar : PubMed/NCBI
3.	Chamberlain G, Fox J, Ashton B and Middleton J: Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing. Stem Cells. 25:2739–2749. 2007. View Article : Google Scholar
4.	Kang SK, Shin IS, Ko MS, Jo JY and Ra JC: Journey of mesenchymal stem cells for homing: strategies to enhance efficacy and safety of stem cell therapy. Stem Cells Int. 2012:3429682012.PubMed/NCBI
5.	Manning BD and Cantley LC: AKT/PKB signaling: navigating downstream. Cell. 129:1261–1274. 2007. View Article : Google Scholar : PubMed/NCBI
6.	Chin YR and Toker A: Akt isoform-specific signaling in breast cancer: uncovering an anti-migratory role for palladin. Cell Adh Migr. 5:211–214. 2011. View Article : Google Scholar : PubMed/NCBI
7.	Worster DT, Schmelzle T, Solimini NL, Lightcap ES, Millard B, Mills GB, Brugge JS and Albeck JG: Akt and ERK control the proliferative response of mammary epithelial cells to the growth factors IGF-1 and EGF through the cell cycle inhibitor p57Kip2. Sci Signal. 5:ra192012.PubMed/NCBI
8.	Kakinuma N, Roy BC, Zhu Y, Wang Y and Kiyama R: Kank regulates RhoA-dependent formation of actin stress fibers and cell migration via 14-3-3 in PI3K-Akt signaling. J Cell Biol. 181:537–549. 2008. View Article : Google Scholar : PubMed/NCBI
9.	Cenni V, Sirri A, Riccio M, Lattanzi G, Santi S, de Pol A, Maraldi NM and Marmiroli S: Targeting of the Akt/PKB kinase to the actin skeleton. Cell Mol Life Sci. 60:2710–2720. 2003. View Article : Google Scholar : PubMed/NCBI
10.	Scita G, Tenca P, Frittoli E, Tocchetti A, Innocenti M, Giardina G and Di Fiore PP: Signaling from Ras to Rac and beyond: not just a matter of GEFs. EMBO J. 19:2393–2398. 2000. View Article : Google Scholar : PubMed/NCBI
11.	Qian Y, Corum L, Meng Q, Blenis J, Zheng JZ, Shi X, Flynn DC and Jiang BH: PI3K induced actin filament remodeling through Akt and p70S6K1: implication of essential role in cell migration. Am J Physiol Cell Physiol. 286:C153–C163. 2004. View Article : Google Scholar : PubMed/NCBI
12.	Enomoto A, Murakami H, Asai N, Morone N, Watanabe T, Kawai K, Murakumo Y, Usukura J, Kaibuchi K and Takahashi M: Akt/PKB regulates actin organization and cell motility via Girdin/APE. Dev Cell. 9:389–402. 2005. View Article : Google Scholar : PubMed/NCBI
13.	Yoeli-Lerner M, Yiu GK, Rabinovitz I, Erhardt P, Jauliac S and Toker A: Akt blocks breast cancer cell motility and invasion through the transcription factor NFAT. Mol Cell. 20:539–550. 2005. View Article : Google Scholar : PubMed/NCBI
14.	Cenni V, Bavelloni A, Beretti F, Tagliavini F, Manzoli L, Lattanzi G, Maraldi NM, Cocco L and Marmiroli S: Ankrd2/ARPP is a novel Akt2 specific substrate and regulates myogenic differentiation upon cellular exposure to H(2)O(2). Mol Biol Cell. 22:2946–2956. 2011. View Article : Google Scholar : PubMed/NCBI
15.	Irie HY, Pearline RV, Grueneberg D, Hsia M, Ravichandran P, Kothari N, Natesan S and Brugge JS: Distinct roles of Akt1 and Akt2 in regulating cell migration and epithelial-mesenchymal transition. J Cell Biol. 171:1023–1034. 2005. View Article : Google Scholar : PubMed/NCBI
16.	Pierini M, Dozza B, Lucarelli E, Tazzari PL, Ricci F, Remondini D, di Bella C, Giannini S and Donati D: Efficient isolation and enrichment of mesenchymal stem cells from bone marrow. Cytotherapy. 14:686–693. 2012. View Article : Google Scholar : PubMed/NCBI
17.	Maraldi T, Riccio M, Resca E, Pisciotta A, La Sala GB, Ferrari A, Bruzzesi G, Motta A, Migliaresi C, Marzona L and De Pol A: Human amniotic fluid stem cells seeded in fibroin scaffold produce in vivo mineralized matrix. Tissue Eng Part A. 17:2833–2843. 2011. View Article : Google Scholar : PubMed/NCBI
18.	Dozza B, Gobbi G, Lucarelli E, Pierini M, Bella CD, Frisoni T, Tazzari PL, Ricci F, Mirandola P, Carubbi C, Giannini S, Donati D and Vitale M: A rapid method for obtaining mesenchymal stem cells and platelets from bone marrow aspirate. J Tissue Eng Regen Med. June 19–2012.(Epub ahead of print). View Article : Google Scholar
19.	Maraldi T, Riccio M, Sena P, Marzona L, Nicoli A, La Marca A, Marmiroli S, Bertacchini J, La Sala G and De Pol A: MATER protein as substrate of PKCepsilon in human cumulus cells. Mol Hum Reprod. 15:499–506. 2009. View Article : Google Scholar : PubMed/NCBI
20.	Cenni V, Sabatelli P, Mattioli E, Marmiroli S, Capanni C, Ognibene A, Squarzoni S, Maraldi NM, Bonne G, Columbaro M, Merlini L and Lattanzi G: Lamin A N-terminal phosphorylation is associated with myoblast activation: impairment in Emery-Dreifuss muscular dystrophy. J Med Genet. 42:214–220. 2005. View Article : Google Scholar : PubMed/NCBI
21.	Duca M, Dozza B, Lucarelli E, Santi S, Di Giorgio A and Barbarella G: Fluorescent labeling of human mesenchymal stem cells by thiophene fluorophores conjugated to a lipophilic carrier. Chem Commun (Camb). 46:7948–7950. 2010. View Article : Google Scholar : PubMed/NCBI
22.	Maraldi T, Bertacchini J, Benincasa M, Guida M, De Pol A, Liotta LA, Petricoin E, Cocco L and Marmiroli S: Reverse-phase protein microarrays (RPPA) as a diagnostic and therapeutic guide in multidrug resistant leukemia. Int J Oncol. 38:427–435. 2011.PubMed/NCBI
23.	Gebäck T, Schulz MM, Koumoutsakos P and Detmar M: TScratch: a novel and simple software tool for automated analysis of monolayer wound healing assays. Biotechniques. 46:265–274. 2009.PubMed/NCBI
24.	Mirandola P, Sponzilli I, Gobbi G, Marmiroli S, Rinaldi L, Binazzi R, Piccari GG, Ramazzotti G, Gaboardi GC, Cocco L and Vitale M: Anticancer agents sensitize osteosarcoma cells to TNF-related apoptosis-inducing ligand downmodulating IAP family proteins. Int J Oncol. 28:127–133. 2006.PubMed/NCBI
25.	Martelli AM, Evangelisti C, Follo MY, Ramazzotti G, Fini M, Giardino R, Manzoli L, McCubrey JA and Cocco L: Targeting the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin signaling network in cancer stem cells. Curr Med Chem. 18:2715–2726. 2011. View Article : Google Scholar : PubMed/NCBI
26.	Wu W-I, Voegtli WC, Sturgis HL, Dizon FP, Vigers GPA and Brandhuber BJ: Crystal structure of human AKT1 with an allosteric inhibitor reveals a new mode of kinase inhibition. PLoS One. 5:e129132010. View Article : Google Scholar : PubMed/NCBI
27.	Chin YR and Toker A: The actin-bundling protein palladin is an Akt1-specific substrate that regulates breast cancer cell migration. Mol Cell. 38:333–344. 2010. View Article : Google Scholar : PubMed/NCBI
28.	Nocentini S, Reginensi D, Garcia S, Carulla P, Moreno-Flores MT, Wandosell F, Trepat X, Bribian A and Del Río JA: Myelin-associated proteins block the migration of olfactory ensheathing cells: an in vitro study using single-cell tracking and traction force microscopy. Cell Mol Life Sci. 2012. View Article : Google Scholar
29.	Fox JM, Chamberlain G, Ashton BA and Middleton J: Recent advances into the understanding of mesenchymal stem cell trafficking. Br J Haematol. 137:491–502. 2007. View Article : Google Scholar : PubMed/NCBI
30.	Stambolic V and Woodgett JR: Functional distinctions of protein kinase B/AKT isoforms defined by their influence on cell migration. Trends Cell Biol. 16:461–466. 2006. View Article : Google Scholar : PubMed/NCBI
31.	Toker A: Achieving specificity in Akt signaling in cancer. Adv Enzyme Regul. 52:78–87. 2012. View Article : Google Scholar : PubMed/NCBI
32.	Virtakoivu R, Pellinen T, Rantala JK, Perälä M and Ivaska J: Distinct roles of AKT isoforms in regulating β1-integrin activity, migration, and invasion in prostate cancer. Mol Biol Cell. 23:3357–3369. 2012.
33.	Moritz A, Li Y, Guo A, Villén J, Wang Y, MacNeill J, Kornhauser J, Sprott K, Zhou J, Possemato A, Ren JM, Hornbeck P, Cantley LC, Gygi SP, Rush J and Comb MJ: AKT-RSK-S6 kinase signaling networks activated by oncogenic receptor tyrosine kinases. Sci Signal. 3:ra642010.PubMed/NCBI