Melanoma in the Eyes of Mechanobiology

cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 1 REVIEW published: 11 Feb ua y 2020 doi: 10.3389/ cell.2020.00054 Edi ed by: Shamik Sen, Indian Ins i u e o Technology Bombay, India Re iewed by: Zhizhan Gu, Dana-Fa be Cance Ins i u e, Uni ed S a es Takashi Ka o, Johns Hopkins Uni e si y, Uni ed S a es *Co espondence: Man ed Radmache [email p o ec ed] †ORCID: Ped o L. G anja o cid.o g/0000-0003-2761-4929 Special y sec ion: This a icle was submi ed o Cell Adhesion and Mig a ion, a sec ion o he jou nal F on ie s in Cell and De elopmen al Biology Recei ed: 15 July 2019 Accep ed: 21 Janua y 2020 Published: 11 Feb ua y 2020 Ci a ion: B ás MM, Radmache M, Sousa SR and G anja PL (2020) Melanoma in he Eyes o Mechanobiology. F on . Cell De . Biol. 8:54. doi: 10.3389/ cell.2020.00054 Melanoma in he Eyes o Mechanobiology M. Manuela B ás1,2,3, Man ed Radmache 4*, Susana R. Sousa1,2,5 and Ped o L. G anja1,2,3† 1Ins i u o de In es igação e Ino ação em Saúde, Uni e sidade do Po o, Po o, Po ugal, 2Ins i u o de Engenha ia Biomédica, Uni e sidade do Po o, Po o, Po ugal, 3Faculdade de Engenha ia, Uni e sidade do Po o, Po o, Po ugal, 4Ins i u e o Biophysics, Uni e si y o B emen, B emen, Ge many, 5Ins i u o Supe io de Engenha ia do Po o, Ins i u o Poli écnico do Po o, Po o, Po ugal Skin is he la ges o gan o he human body wi h se e al impo an unc ions ha can be impai ed by inju y, gene ic o ch onic diseases. Among all skin diseases, melanoma is one o he mos se e e, which can lead o dea h, due o me as iza ion. Mechano ansduc ion has a c ucial ole o mo ili y, in asion, adhesion and me as iza ion p ocesses, since i deals wi h he esponse o cells o physical o ces. Signaling pa hways a e impo an o unde s and how physical cues p oduced o media ed by he Ex acellula Ma ix (ECM), a ec heal hy and umo cells. Du ing hese p ocesses, se e al molecules in he nucleus and cy oplasm a e ac i a ed. Melanocy es, ke a inocy es, ib oblas s and he ECM, play a c ucial ole in melanoma o ma ion. This manusc ip will add ess he syne gy among melanocy es, ke a inocy es, ib oblas s cells and he ECM conside ing hei mechanical con ibu ion and ele ance in his disease. Mechanical p ope ies o melanoma cells can also be in luenced by pigmen a ion, which can be associa ed wi h changes in s i ness. Mechanical changes can be ela ed wi h he adhesion, mig a ion, o in asi eness po en ial o melanoma cells p omo ing a high me as iza ion capaci y o his cance . Mechanosensing, mechano ansduc ion, and mechano esponse will be highligh ed wi h espec o he mo ili y, in asion, adhesion and me as iza ion in melanoma cance . Keywo ds: melanoma, mechanobiology, melanocy es, ke a inocy es, in asion, adhesion, mig a ion, me as iza ion MELANOMA Melanoma is one o he mos se e e skin cance s in humans wi h a mo ali y a e o 80% (Ho sch oe e al., 2017), due o he high esis ance o his umo o adio he apy and chemo he apy (Uong and Zon, 2010). The esis ance o hose ea men s is due o high numbe o mu a ions (app ox. 75%) a ec ing speci ic genes, which lead o an inc easing p oli e a ion and su i al (Koza e al., 2019). Immuno he apy is being exploi ed and in es iga ed, and can be g ouped in i e ypes: (i) a ge ed an ibodies (Sanlo enzo e al., 2014;He zbe g and Fishe , 2016); (ii) adop i e cell he apy (Sanlo enzo e al., 2014); (iii) oncoly ic i us he apy (F anklin e al., 2017); (i ) cance accines (Tue enbe g e al., 2007;Rod iguez-Ce dei a e al., 2017); and ( ) immunomodula o s (Johnson e al., 2014;Fa ies, 2016;Cance Resea ch Ins i u e, 2019). Chemo he apy and immuno he apy, can be combined (biochemo he apy) ega ding he s age disease (Sanlo enzo e al., 2014;Koza e al., 2019). The disease appea s due o he malignan ans o ma ion o melanocy es loca ed in F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 1Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 2 B ás e al. Melanoma in he Eyes o Mechanobiology he s a um basale o he epide mis al hough i is no clea how a melanocy e becomes malignan (Uong and Zon, 2010). Banda chi e al. (2010) sugges ed ha a combina ion o up and down egula ion o ac o s in se e al molecula pa hways, seem o be he mechanism o ans o ma ion o no mal in o malignan melanocy es. I he disease is de ec ed in an ea ly s age, he e is a high p obabili y o disease con ol, howe e , when me as iza ion occu s, he 5 yea s li e expec ancy d ops o 14%. Some epidemiological e idence ela ed o melanoma incidences in whi e and da k skinned popula ion (Banda chi e al., 2013) was p esen ed by he au ho s whe eas B ad o d (2009) p esen ed epidemiological da a abou he o al numbe s o melanoma incidences in he wo ld. Da k skin esul s in a lowe incidence o skin cance due o he p o ec ion o he inc eased epide mal melanin, which il e s UV ligh wice as good as whi e skin o Caucasian people (B ad o d, 2009). Malignan melanoma a e diagnosed by he ABCD ule, whe e A s ands o asymme y, B o bo de s i egula i y, C o colo a ia ion and D o diame e highe han 6 mm (Banda chi e al., 2010). Melanoma shows h ee pe cep ible s eps in umo p og ession: (i) one only a ec ing he epide mis; (ii) a second one a ec ing bo h he epide mis and he supe icial papilla y o he de mis; and (iii) a hi d one a ec ing he epide mis, de mis and hypode mis ( e ical g ow h melanoma) (Banda chi e al., 2010). Haass e al. (2005) di ided he p ocess o melanoma de elopmen in he ollowing s eps: (i) he damage o adhesion be ween ke a inocy es by down- egula ion o E-cadhe in, P-cadhe in, desmoglein, and connexins, which is in luenced by g ow h ac o s p oduced by ib oblas s o ke a inocy es; (ii) he in e ac ion o melanoma wi h ib oblas s and/o melanoma wi h melanoma cells no mally no ound in melanocy es, is up- egula ed by MCAM and N-cadhe in; and (iii) al e a ion o in eg in exp ession, inducing he loss o basemen memb ane ancho age (Haass e al., 2005). Acco ding o he A Melanoma Founda ion (AIM), melanoma de elops in a simila way han o he cance s: he DNA o a gene ha con ols cell di ision and p oli e a ion is mu a ed. This damaged gene esul s in a lack o con ol o cell di ision and g ow h. This DNA mu a ion is passed o he daugh e cell du ing di ision. Melanoma is o igina ed om a melanocy e ha expe ienced oo many mu a ions g owing in an abno mal way. In he case o melanoma, his DNA mu a ion is caused by an o e exposu e o UV adia ion, a ec ing he melanocy es (A Melanoma Founda ion, 2014). When he e is DNA damage, se e al genes may be a ec ed, esul ing in encoding e o s o se e al molecules hampe ing hei unc ion (A Melanoma Founda ion, 2014;Kunz, 2014) and he e is a isk o a melanocy e o gene a e a melanoma. I he melanoma is no supp essed o ea ed, i will sp ead along he epide mis, a e pene a ing he de mis and hypode mis, ge ing in con ac wi h he lymph and he blood essels (A Melanoma Founda ion, 2014). The main unc ion o melanocy es is o p oduce melanin ( esponsible o skin pigmen a ion) ha abso b UV ligh o a oid ke a inocy es’ DNA damage (Hi obe, 2014). Ke a inocy es will dis ibu e melanin o he uppe pa s o he skin laye (Wang e al., 2017). Wang e al. (2017) also men ioned ha ho monal ac o s, amily his o y and cosme ics a e he h ee main igge ing and agg a a ing ac o s o melanoma de elopmen . When a p ima y melanoma is o med, i ac i a es some signals, which will p o ide condi ions ha make he p ima y umo s ong such as: (i) a achmen o he wall o he blood and/o lympha ic essels, enabling he mo emen h ough a new o gan; (ii) enough nu ien s o g ow; and (iii) i is esis an o he immune sys em. Wi h he h ee condi ions men ioned be o e he p ima y umo has he capaci y o p omo e he me as iza ion (A Melanoma Founda ion, 2014). Why can melanoma de elop in some body pa s ha a e no exposed o sunligh ? I was sugges ed ha he e a e di e en changes occu ing in genes o issue cells exposed, compa ed wi h he changes o genes o issue cells no exposed o UV ligh . These mu a ions a e ansmi ed in o he nex gene a ions. Some genes appea du ing melanoma de elopmen , being no gene ic. The ans o ma ion o melanocy es in o melanoma cells is a e y complex p ocess, which in ol es se e al signaling pa hways e iewed by Paluncic e al. (2016). B ie ly, melanocy es a e o med om hei p ecu so s: melanoblas s. Melanoblas s di e en ia e in o melanocy es, which ge ma u e and s a melanin p oduc ion on melanosomes, which will be ans e ed o ke a inocy es. Melanomagenesis depends on he mic oen i onmen , gene ic and en i onmen al ac o s. Al hough gene ic ac o s a e c ucial, hey a e no su icien o induce melanomagenesis (Paluncic e al., 2016). Melanoma umo s a e di e en om pa ien o pa ien . The en i onmen , whe e melanoma umo s g ow, is e y complex depending on he in e ac ions wi h ECM, mic o ascula u e, ib oblas ic cells and changing concen a ion o g ow h ac o , cy okines, and nu ien s like glucose, oxygen, e c (Paluncic e al., 2016). Some signaling pa hways a e mo e in ol ed in umo de elopmen , whe eas o he signals a e mo e c ucial in he me as iza ion p ocess. The desc ip ion o biochemical pa hways signaling is beyond o his e iew, howe e he eade s can ind mo e in o ma ion in di e en sou ces (Dissanayake e al., 2007;S aussman e al., 2012;Mo iceau e al., 2015;S a k e al., 2015;Wang e al., 2016; Ahmed and Haass, 2018;Kode e al., 2018). Melanoma can be pigmen ed o no pigmen ed. The colo o melanoma is due o he exis ence o melanin, sec e ed and s o ed in melanosomes in he cy oplasm o melanocy es. The e a e wo ypes o melanin: eumelanin (has a black colo ) and pheomelanin (has a ed/yellow colo ). No mal melanocy es may o may no sec e ei he ype o melanin (Lassalle e al., 2003). Mechanical p ope ies and cell pigmen a ion a e co ela ed (Sa na e al., 2013, 2014). As melanoma needs a high amoun o i on and coppe , he de elopmen o subs ances ha a ge hese ions could be ano he me hodology (Go ode sky e al., 1986). MECHANOBIOLOGY OF SKIN CELLS Cu en ly, mechanobiology b ings oge he biologis s and biophysicis s in o de o be e unde s and he ole o mechanical o ces in cell mo ili y, adhesion and in asion. Fo de eloping he apies ha a oid he in asi eness o cance cells, i is c ucial o unde s and he mechanisms o umo in asion. The mechanical s i ness o umo s is co ela ed wi h i s in asi eness, which was demons a ed by a s udy wi h cance o a ian cells: mo e in asion co esponds o so e cells esul ing in de o ma ion and shape F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 2Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 3 B ás e al. Melanoma in he Eyes o Mechanobiology changes, being sui able o me as iza ion (Swamina han e al., 2011). Makale (2007) aised h ee hypo heses co ela ing cance and cell mechanobiology: (i) he cell biology o umo in asion is a key o unde s and cance ea men ; (ii) cance cells ac i a e physiological mechanisms and mig a o y p ocesses ha a e used by cells usually du ing emb yo mo phogenesis; and (iii) o ces ha a e sensed by he issue and cells igge he di e en ia ion and sp eading o umo s. In ac , mechanical o ces a e gene a ed by issues, being sensed by cells, du ing emb yogenesis and umo expansion (Makale, 2007). In melanoma he Epi helial o Mesenchymal T ansi ion (EMT) swi ching, and o ces behind his p ocess (Wei e al., 2015) a e c ucial p ocesses ega ding he loss o connec ions be ween melanocy e and ke a inocy es; melanocy es acqui ed he emb yonic pheno ype, p omo ing cells in asion, hei in a asa ion in body luids and he ex a asa ion o o he o gans. A i ing a hei a ge des ina ion, umo cells a e e e ed o hei o iginal pheno ype known as Mesenchymal o Epi helial T ansi ion (MET), es ablishing seconda y umo s. The p oduc ion o me allop o einases (MMP) will induce ECM deg ada ion, a o ing me as iza ion (Paluncic e al., 2016). Mig a ion o collec i e cells in e ac wi h he ex acellula en i onmen ia in eg ins (Hege eld e al., 2002). When no mal cells a e comp essed because o a umo , hese o ces will be sensed by cell body, h ough memb ane molecules, he in eg ins, which a e connec ed o he cy oskele on, and o he nucleus. All cells espond o hese ex e nal o ces o ganizing cellula componen s in he cell and gene a ing biochemical signals ha will help hem o adap o ex e nal physical p essu e. The no mal ECM p oduced by ib oblas s also induces p essu e on umo cells ia i s cy oskele on, esul ing in cell and nucleus de o ma ion. A consequence o nucleus comp ession could be he modula ion o gene exp ession ha may induce changes in cell iabili y, p omo ing locomo ion, due o he adop ion o a mo e malignan pheno ype. In spi e o high umo s oma s i ness, umo cells a e de o mable and ha e some deg ee o plas ici y (Makale, 2007). In melanoma, he ela ion among melanocy es, ke a inocy es, ib oblas s and he ECM media e and con ol his disease. In he nex pa ag aphs, mechanobiology o hese cell ypes and ECM con ibu ion o melanoma will be add essed. The Role o Melanocy es Melanocy es a e loca ed in he basal laye o he epide mis, eyes, ea s, hai and meninges (Banda chi e al., 2013). Melanocy es ha e a dend i ic shape (Figu e 1). They a e o igina ed om mig a o y emb yonic cells, called neu al c es cells, which ha e a high capaci y o mig a ion (Uong and Zon, 2010). Melanocy es a e he cells esponsible o skin pigmen a ion, h ough he p oduc ion o melanin, also named melanogenesis (Lassalle e al., 2003). They a e p esen in he epide mis and exhibi long dend i ic p o usions, which a e ex ended h ough he epide mal ke a inocy es. Melanocy es syn hesize melanosomes and anspo hem o he ke a inocy es (Zhang e al., 2004). Melanocy es adhe e o ke a inocy es h ough E-cadhe ins, Desmoglein (DSg)/Desmocollin (DSc), Connexins (CX) molecules (Li e al., 2003) and o he basemen memb ane h ough in eg ins (Wang e al., 2016). The a io o melanocy es and ke a inocy es is a ound 1:36/1:40. 8 o 12% o malignan melanoma a e associa ed wi h mu a ions on CDKN2A (cyclin- dependen kinase Inhibi o 2) gene (Banda chi e al., 2013). Uong and Zon (2010) also men ioned ha he pa hways in ol ed in melanocy e de elopmen and p oli e a ion s a o be sub e ed in melanoma o ma ion. BRAF and NRAS (a gene o he RAS amily iden i ied o he i s ime a human neu oblas oma) (Pugh e al., 2013) o e ac i e mu a ions a e ound in 65 and 20% o melanomas, espec i ely. CDKN2A is lacking by melanoma cells, consequen ly he encoding supp esso genes (INK4a and ARF) a e no ac i e (Banda chi e al., 2010). Malignan melanocy es a e cha ac e ized by changes o biophysical p ope ies (like shape, elas ici y and s i ness) which will con ibu e o he cell locomo ion, in asion and me as iza ion in o he o gans. In his case, he plas ici y o melanocy es is e y impo an , since hey need o be highly de o mable. Since he mechanical p ope ies o melanoma cells may be a ec ed by he p oduc ion o melanin, his has o be conside ed (Sa na e al., 2013). Melanin and melanogenesis may ac in wo di e en di ec ions: melanin ac as a p o ec ion shield o adia ion, howe e i also a enua es he e ec i eness o adio he apy, chemo he apy and pho o he apy in he case o melano ic melanoma. This cha ac e is ic can p o ide umo g ow h and p og ession (Slominski e al., 2015). AFM was used o obse e he shape o human epide mis melanocy es in cul u e and o obse e he melanosome ans e in i o. Filopodia o igina ed om he dend i e ips o he cell body o melanocy es, which con ained melanosomes ha e been obse ed (Zhang e al., 2004). Van Den Bossche e al. (2006) hypo hesized di e en mechanisms o melanin ans e : (i) ke a inocy es phagocy e he melanocy e dend i es; (ii) he melanosomes elease melanin in o he ex acellula space by melanosome exocy osis, being in e nalized by he ke a inocy es; (iii) melanosomes a e anspo ed ia memb ane nano ubes; and (i ) he melanocy es en iched o melanosomes a e phagocy osed by he ke a inocy es (Van Den Bossche e al., 2006). Wu e al. (2012) s udied melanocy es o mice lacking myosin Va (Myo5a). This p o ein is in ol ed in he accumula ion o melanosomes in he melanocy e dend i es. When myosin Va is p esen , melanosomes s ay concen a ed in he cen e o cance ous melanocy es educing melanosome ans e o ke a inocy es (Wu e al., 2012). Wäs e e al. (2016) s udied he ela ion be ween UV skin adia ion and skin pigmen a ion esponse. They concluded ha when pigmen ed skin is exposed o UVA, he e is a spillage o ex acellula esicles om he melanocy e’s plasma memb ane. This does no happen when pigmen ed skin is exposed o UVB (Wäs e e al., 2016). AFM was used o s udy he mo phological changes o melanocy es exposed o he melanocy e s imula ed ho mone α-MSH, which induces melaniza ion (Shin e al., 2014). The au ho s ound ha he oughness inside he po e-like s uc u es on dend i es inc eased and MSH p oduced an inc ease in he size and densi y o he melanosomes. Howe e , ega ding he esponse o he p esence o α-MSH, mo e s udies a e needed o co ela e mo phological shape al e a ions wi h he mo emen o melanosomes (Shin e al., 2014). Human melanocy e’s pigmen a ion in melanoma cells a e also impo an in egula ion F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 3Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 4 B ás e al. Melanoma in he Eyes o Mechanobiology FIGURE 1 | Schema ic ep esen a ion o skin and cell popula ion. Melanocy es a e loca ed in he basal laye o epide mis. Each melanocy e is connec ed o se e al ke a inocy es. Melanosomes (black spo s) a e dis ibu ed in he den i es, p oduce and s o e melanin. The dend i es allow he ans e o he melanin o ke a inocy es, which will be ans e ed o he s a um co neum laye o he epide mis. o elas ic p ope ies acco ding o Choi e al. (2014). The au ho s s udied he e ec o subs a es wi h di e en s i ness on he pigmen a ion, using no mal human melanocy es (NHM) and melanoma cells (MNT1) cul u ed on polydime hylsiloxane (PDMS) coa ed wi h laminin. They obse ed a educ ion o dend i e o ma ion in bo h cell ype as well as a di e ence on he ans e ence a e o melanosomes o bo h cell ype on so e subs a es. Cells cul u ed on s i e subs a es showed lowe mig a ion capaci y once ac ion o ces o ma ion a e mo e e iden h ough ia ocal adhesion in eg in complexes (Choi e al., 2014). Based on some s udies o cell elas ici y, cance cells ypically show a dec ease in s i ness ela i ely o no mal cells. Sa na e al. (2013) desc ibed a s udy o elas ic p ope ies compa ing melanoma cells and heal hy melanocy es. Basically, he au ho s used AFM o de e mine he s i ness o human melanoma cells SKMEL-188 wi h di e en s ages o pigmen a ion and compa ed wi h SKMEL-188 cells no pigmen ed and wi h NHMs heal hy human melanocy es (Sa na e al., 2013). They ound ha heal hy melanocy es we e so e han cance cells. This ea u e could be ela ed wi h he amoun o melanin p esen in melanoma cells, esul ing in some limi a ions o cance diagnosis using he s i ness. Lazo a and Pawelek (2009) men ioned ha melanoma cells do no exc e e he pigmen like heal hy melanocy es do, which may esul in pigmen accumula ion in melanoma cells in ela ion o melanocy es. This could explain he highe s i ness o cance cells, since hey a e o en highly pigmen ed (Lazo a and Pawelek, 2009). In conclusion, by AFM nanoinden a ion assays i is no possible o dis inguish melanoma cells om melanocy es. AFM can be used as a complemen a y echnique, howe e , his s udy was use ul in unde s anding be e pigmen ed cell elas ici y. Sa na e al. (2014) also men ioned he need o unde s and i he e is a co ela ion be ween he pigmen a ion o melanoma cells and he me as iza ion pheno ype p ocess. In his p ocess, malignan cells mus pass he epi helial ba ie , ha ing he capaci y o de o m hei cell body, including he nucleus o mig a e h ough issue. Cells wi h low Young’s moduli exhibi a g ea po en ial o in ade issue (Sa na e al., 2014), so his is why biophysical p ope ies, as cell elas ici y, mus be aken in o accoun . The au ho s showed ha melanin g anules inhibi he ansmig a ion po en iali ies o melanoma cells. Fo non-pigmen ed melanoma cells, he s i ness was lowe , and he ansmig a ion e iciency was highe . This s udy con ibu ed wi h new insigh s abou he ela ionship be ween pigmen a ion and elas ici y o melanoma cells in e ms o mig a ion and in asion. Some subs ances a e esponsible o he e ac ion o melanocy e dend i es. I o e al. (2006) ha e s udied he e ec o dend i e e ac ion using a la onoid subs ance. In his case, TYRP-1 signaling pa hway is in e up ed, once his molecule is impo an o melanosome mig a ion. Rho signaling molecule, when ac i a ed induces dend i ic e ac ion, educing he melanosome ans e o he ke a inocy es bu does no a ec he amoun o melanin p oduc ion. Howe e , his melanin will no be ans e ed due o he dend i e’s e ac ion. The mechanism o dend i e e ac ion ia RhoA in ol es changes in cy oskele on and mic o ubules o ganiza ion due o ac in and ubulin-βdisassembly. These changes will induce s ess ibe o ma ion p omo ing ac in polyme iza ion and eo ganiza ion o mic o ubules (I o e al., 2006). Rac is in ol ed in melanocy e dend i e g owing, inducing he appea ing o lamellipodia (Sco , 2002). The dend i e ips o melanocy es a e also s imula ed by he α-MSH and endo helin – 1 p o eins om ke a inocy es ha a e su ounding he melanocy es and he ECM p oduced by he ke a inocy e (Kippenbe ge e al., 1998). Kippenbe ge e al. (1998) demons a ed ha co-cul u es o melanocy es and ke a inocy es in he p esence o high concen a ion o calcium, showed shee s o ke a inocy es o ma ion, su ounding by melanocy es wi h pola dend i es ex ended a ound he di e en ia ed ke a inocy es. Di e en ia ed ke a inocy es will ha e s able a eas o con ac wi h melanocy es due o he calcium g adien . The no mal melanocy es ha lose hei dend i es soon will be in an apop osis s a e (Kippenbe ge e al., 1998). The senescence o p ima y melanocy es could also be impo an in he melanoma appea ance (Feue e e al., 2019), howe e , he e a e no s udies ha co ela e melanocy es’ senescence wi h mechanical p ope ies measu ed by AFM. Some s udies showed F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 4Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 5 B ás e al. Melanoma in he Eyes o Mechanobiology a co ela ion be ween he melanocy es aging and senescence bioma ke s (Wang and D eesen, 2018) and exposi ion o melanocy es o ligh (Benne e al., 2017). AFM is a powe ul ool o assess he mo phological and elas ic p ope ies ega ding he in e ace be ween p ima y melanocy es and p ima y melanoma wi h and wi hou senescence. In opposi ion o he melanoma cell lines ha a e immo alized and hus, submi ed o se e al passages, he p ima y melanocy es and p ima y melanoma cells should be assessed in o de o e alua e wha passage he senescence will no in e e e wi h mechanical p ope ies. The quan i ica ion o senescence le el can be pe o med using he quan i ica ion o he Popula ion Double Le el (PDL) me hod (K use and Pa e son, 1973). The Role o Ke a inocy es Ke a inocy es a e he p edominan cell ype in he epide mis, mos o hem (90%) being loca ed a he ou mos epide mis laye (McG a h e al., 2004). Ke a inocy es a e connec ed o each o he h ough ac in cell junc ions (Hsu e al., 2018). The main unc ion o ke a inocy es is o o m a ba ie agains pa hogenic mic oo ganisms, hea , UV adia ion and wa e loss. They also p oduce cy okines ha a ac leukocy es o he loca ion o pa hogen in asion (Hall e al., 2014). Ke a inocy es also p oduce di e en ke a ins. Ke a ins ( ype 1, 5, 10, and 14) a e he main s uc u al p o ein o hei cy oskele on, all o which play an impo an ole in he mechanics o hese cells (Ramms e al., 2013). Ke a ins become al e ed when esponding o s esses, esul ing in i s modi ica ion and eo ganiza ion (Ka an za, 2011). Ke a inocy es become co neocy es, when hey a e comple ely di e en ia ed, losing hei nucleus and cy oplasmic o ganelles (Mi, 2009). Di e en ia ion o ke a inocy es is d i en by a calcium g adien , which is es ablished be ween he in e io o he cell and he ECM (P oksch e al., 2008) and egula ed by i amin D3 (Bikle, 2004), ca hepsin E (Kawakubo e al., 2011), TALE homeodomain ansc ip ion ac o s (Jackson e al., 2011), and hyd oco isone (Rheinwald and G een, 1975). Ke a inocy es a e a ec ed by senescence, losing hei biological unc ion wi h age (O ioli and Dellamb a, 2018). Ke a in, ac in ilamen s and mic o ubules o m he cy oskele on ne wo k o ke a inocy es, p o iding shape and s uc u e o cells allowing ansmission o mechanical loads (Sil e and Sipe ko, 2003). Fo ke a inocy es mig a ion one o he mos impo an pa ame e s is he subs a e s i ness (Ramms e al., 2013), since i is expec ed ha he ke a inocy es mus change hei shape du ing mig a ion h ough he epide mis un il hey each he ou mos laye and become co neocy es (Za koob e al., 2015). This change in shape will a ec also he componen s exis ing inside he cy oplasm, namely he cy oskele on, in which ke a in ibe s a e c ucial (Ramms e al., 2013). Ramms e al. (2013) used AFM o p obe ke a inocy es (in he nuclea egion and in he cell body) de icien in ke a ins inding ha hey a e so e han heal hy ones. In addi ion, wi h magne ic weeze s, he iscous con ibu ion o he displacemen o magne ic beads inc eased, co obo a ing he esul s achie ed by AFM. These au ho s also ound ha he so ening was e e sible and could depend on he e-exp ession o wo ypes o Ke a ins (K4 and K15) only (Ramms e al., 2013). Ba e al. (2014) epo ed he skin agili y in mice and hampe ed desmosomes adhesion when he e is a lack o all ke a ins Ba e al., 2014. AFM showed a educ ion o adhesi e o ces and memb ane s abili y in mu ine ke a inocy es when lacking ke a in ilamen s (Vielmu h e al., 2018). Adhe en junc ions (AJ) ha e unc ions in mechanosensing and ansducing mechanical o ces be ween he plasma memb ane and he ac omyosin cy oskele on. Desmosomes and in e media e ilamen s p omo e he mechanical s abili y o main ain issue a chi ec u e and in eg i y when hey a e submi ed o mechanical s ess. Bo h (AJ and desmosomes) connec he ac in and ke a in ilamen ne wo k o adjacen cells. Desmosomes a e impo an o in e cellula cohesion, whe e ke a ins de e mine cell mechanics bu a e no in ol ed in ension gene a ion (Ha z eld e al., 2017). Za koob e al. (2015) in es iga ed he ole o subs a e s i ness on ke a inocy e colony o ma ion (in i o) du ing epi helial o ma ion, due o calcium exchange du ing ke a inocy e cell cul u e. Ke a inocy es cul u ed on so polyac ylamide subs a es (E= 1.2 KPa) p esen ed small con ac a eas, an inc ease in mig a ion eloci y and in colony o ma ion compa ed o hose cul u ed on s i polyac ylamide gels (E= 24 KPa) (Za koob e al., 2015). How melanoma cells escape he ke a inocy e’s con ol in ading o he issues and o m me as asis, is s ill unclea . In a heal hy issue, he e is a homeos asis main ained by each cell. In his no mal issue, ke a inocy es a e he “su eyo s” o melanocy es, con olling hei g ow h and beha io h ough complex pa ac ine g ow h ac o s and cell-cell adhesion molecules. When al e a ions occu o his homeos a ic balance, cell-cell adhesion and cell communica ion molecules change and induce melanoma de elopmen esul ing in he damage o he epide mal melanin uni , inducing he con inuous melanocy e p oli e a ion, and e en ually leading o melanoma de elopmen (Haass e al., 2005). Chung e al. (2011) sugges ed ke a inocy es-de i ed ECM ac o s may ac as egula o s o melanocy es. The au ho s showed ha laminin-332, a componen o basemen memb ane, plays a c ucial ole in he adhesion and mig a ion o melanocy es and melanoma. I is known ha ke a inocy es egula e he beha io o melanocy es such as p oli e a ion, melanin syn hesis and dend i e o ma ion and he ECM also egula es di e en cell beha io s (Chung e al., 2011). The Role o Fib oblas s and he ECM Fib oblas s a e p oduced in he bone ma ow. They syn hesize he componen s o he ECM, o ins ance collagen, glycosaminoglycans and elas ic ibe s. Fib oblas s also p oduce a s uc u al ne wo k called “s oma” in animal issues (connec i e issue), playing an impo an ole in cance and in wound healing (Desja dins-Pa k e al., 2018). Kwa e al. (2019) men ioned ha cance is associa ed wi h ib oblas s and how hese ib oblas s a ec me as iza ion. Fib oblas s p oduce high amoun s o ECM molecules, cy okines and g ow h ac o s con ibu ing o he s oma, cance p og ession and me as iza ion due o se e al pa hways. These pa hways could be a a ge o new cance ea men s a egies o be de eloped. Howe e , i is di icul o cha ac e ize his sub ype o ib oblas s and o iden i y speci ic molecules in ol ed in his p ocess (Kwa e al., 2019). Mechanosensing has a la ge impac in Cance -Associa ed Fib oblas s (CAFs). Fib oblas s sense F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 5Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 6 B ás e al. Melanoma in he Eyes o Mechanobiology mechanical s esses and ac i a e mechanosignaling p ocesses, which use mechano ecep o s loca ed in he cell memb ane, he cy oskele on and ansc ip ion ac o s. The ECM s i ness is de e minan in he mechanosignaling, which by i sel can ac i a e he mechanosensing pa hways in CAFs, leading o ECM p oduc ion and s i ness inc easing, suppo ing di e en CAFs di e en ia ion (Kwa e al., 2019). The s udy o mechanosignaling and ib oblas s esponse o he ECM s i ness was conduc ed using hyd ogels, o mimic pa hologically he ECM o issues. Viji Babu e al. (2018) ha e used he AFM o s udy he s i ness and iscoelas ici y o no mal, sca and Dupuy en’s disease ib oblas s, g owing on op o so (1 KPa) and s i (50 KPa) hyd ogels. Basically, a MLCT- Bio py amidal can ile e (nominal sp ing cons an o 0.01 N/m, B uke , Uni ed S a es) was used o inden he ib oblas s, using he z-s ep esponse me hodology, in which he elas ici y and iscosi y was assessed (Figu e 2). The au ho s wan ed o e alua e he e ec o TGF-αin he ib oblas ’s elas ici y. The au ho s concluded ha Dupuy en’s ib oblas s inc eased hei elas ic moduli and became s i e in TGF-β1 p esence, whe eas hey did no ind signi ican changes in he elas ic moduli o sca and no mal ib oblas s, be o e and a e addi ion o TGF- β1. Dupuy en’s ib oblas s p esen ed a wide numbe o well- o ganized s ess ibe s and bundles o s ess ibe s we e e en hicke in p esence o TGF-β1 (Viji Babu e al., 2018). RhoA signaling is he key o he ib oblas ’s eac ion o ECM s i ness. RhoA is ac i a ed by a s i ma ix, causing cell con ac ion and ac in polyme iza ion, inducing he di e en ia ion o ib oblas s in myo ib oblas s. The e o e, CAFs need he ac i a ion o his pa hway, since i is equi ed o hei con ac ile and in asi e p ope ies. I has been men ioned ha he e a e se e al CAF sub ypes, howe e , he e is a lack o in o ma ion abou hei plas ici y, which could be impo an o he p og ession and me as iza ion o cance (Kwa e al., 2019). AFM could gi e new insigh s ega ding he cha ac e iza ion o ib oblas s sub ype based in hei mo phome ic cha ac e is ics (shape ac o , olume and a ea) and mechanical p ope ies. As a as we know, he e is no any speci ic s udy in li e a u e, which FIGURE 2 | Basic componen s o (A) AFM and (B) o ce cu e wi h (C) iscoelas ic c eep esponse measu emen . (A) The basic ou componen s o AFM (i) a lase diode, (ii) a can ile e o 0.01 N/m sp ing cons an wi h 30 nm adius AFM py amidal ip, (iii) a posi ion-sensi i e pho o de ec o (PSPD), and (i ) xyz-piezo s age. (B) Sample inden a ion by he AFM ip ob ains he o ce cu e ha gi es he app oach ( ed a ow) and e ac (blue a ow) cu e on de lec ion s. Z-heigh g aph, and appa en Young modulus was calcula ed by applying He z model o he app oach cu e. In c eep esponse cu e (C), he Z-heigh p o ile (i) shows he app oach and e ac amp owa d he cell o 3 s, and in-be ween, he e is a z s ep, which is applied a = 1.5, which is enla ged in panel (ii).(iii) The de lec ion da a show global c eep o he cell, which includes he c eep a e loading and unloading s ep, which is enla ged in panel (i ), and global c eep was de e mined by he exponen ial i (black cu e) and was sub ac ed o quali a i e analysis ( ep oduced wi h pe mission om John Wiley and Sons wi h Copy igh Clea ance Cen e ’s Righ , Jou nal o Molecula Recogni ion, 2018) (Viji Babu e al., 2018). F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 6Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 7 B ás e al. Melanoma in he Eyes o Mechanobiology could be use ul o he de elopmen o he apies di ec ed o a speci ic ib oblas sub ype ela ed wi h melanoma. This s udy could also p omo e new insigh s o de elop he apeu ic s a egies o o he cance ypes. The ECM is composed by (i) he s uc u al p o eins collagen and elas in; (ii) adhesion p o eins such as ib illin, ib onec in, i onec in and laminin; and (iii) he p o eoglycans (which ha e a p o ein co e bound o glycosaminoglycans) (Makale, 2007). The ECM is esponsible o cellula suppo . Va ia ions in i s s uc u e and composi ion al e he mechanical p ope ies o he ECM. Gene ally, he ex e nal o ces applied o cell-ma ix complexes a e ansmi ed o he in e io o he cell by in eg ins acco ding o h ee s eps (Roca-Cusachs e al., 2009): (i) in eg ins binding o ECM molecules; (ii) ansmission o o ces o he in e io o he cells, h ough in eg ins, whe e hey a e ans o med in o biochemical signals (mechano ansduc ion); and (iii) in eg ins binding o cy oskele on, ansmi ing he o ces all o e he cell, and ein o cing hei adhesion p ope ies o esis he o ce. The in eg ins inside o cell ec ui se e al o he molecules, which bind o he cy oskele on and/o signaling p o eins o o m wha is called “ ocal adhesion” o esis o he ex e nal o ce (Roca-Cusachs e al., 2009). Wha a e he mechanical cues sensed by cells when o ces a e applied om he ou side? In eg in-associa ed complexes (IACs) in cellula memb anes es ablish he connec ion be ween he ECM and he cy oskele on o he cell, ia ac in ilamen s (Haase e al., 2016). Haase e al. (2016) ound ha physical o ces s emming om he ECM ha e a huge impac on gene egula ion; i has been hypo hesized ha gene exp ession is a ec ed by nucleus de o ma ion, which was obse ed in 3T3 ib oblas s. The ac in and mic o ubule ne wo k gene a es nuclea de o ma ion. The de o ma ion o he nucleus will be aniso opic being obse ed along he sho e axis and is in insic o he nucleus. This beha io is due o he ch oma in o ganiza ion and lamin-A exp ession (Haase e al., 2016). The ECM uses he concep o “ enseg i y,” which employs ensile o ces and igid suppo s uc u es o dis ibu e and manage he loads media ed by he cellula cy oskele on complex. In he cy oskele on mic o ilamen s and mic o ubules, which a e connec ed o he ECM, he e a e o ces in be ween (Makale, 2007). This means ha he p essu e exe ed on he ECM will in luence he cell’s beha io in e ms o di e en ia ion, mo ili y, adhesion, in asion, and me as asis (Makale, 2007). ECM mechanical p ope ies a e di ec ly ela ed wi h elas in ibe s, collagens, glycosaminoglycans (GAGs) and p o eoglycans. Elas in ibe s con ibu e o cyclic s e ching du ing li e; collagen ibe s con ibu e o he issue s i ness and mechanical s eng h o issues and GAGs a ec ECM iscoelas ici y, o ming b idges be ween he collagen ibe s and con ibu ing o he comp essi e issue s i ness. Cells sense he o ces exe ed by he ECM, con e ing hese s imuli in o in acellula signal pa hway, which u he downs eam egula e ansc ip ional changes (Hsu e al., 2018). Se e al mechanosenso s a e iden i ied in cells: glycocalyx, lipid a /ca eolin-1, cell adhesion s uc u es (in eg in, hemi- desmosomes and ocal adhesion), ion channels (calcium-sensing ecep o ), ansien ecep o po en ial channels, connexins, in e cellula complexes (desmosomes o cadhe in) and he cy oskele on (pa icula ly he ac in ilamen s) (Hsu e al., 2018). Any abno mali y ha causes changes in he ECM mechanical p ope ies, cell in e ac ions, o cellula esponse o mechanical s imuli will induce ensional homeos asis, con ibu ing o se e al diseases like cance and ib osis (Hsu e al., 2018). In eg ins ( ansmemb ane p o eins) a e esponsible o ansmission o o ces be ween ECM and he in e io o he cell. Tumo cells exp essed he a b3 in eg in and he in asi eness o melanoma cells depends on he exp ession o β-3 and β-1 in eg ins, which a e ela ed wi h hei agg essi e p ope ies (Pe i cle c e al., 1999;Hege eld e al., 2002). The e o e, one he apeu ic s a egy could con ibu e o he educ ion o hese in eg ins exp ession (Makale, 2007). S i ness o umo s is impo an since i ac i a es biochemical pa hways ega ding he cell cycle, EMT, cell mo ili y and leads o comp ession in neighbo ing heal hy cells. These heal hy cells may hen ac i a e umou ogenic pa hways (B ode s-Bondon e al., 2018). Cell sp eading depends on subs a e s i ness, which, in u n, depends on he complex adhesion ype. A7 melanoma cells we e seeded in polyac ylamide gels lamina ed wi h collagen ype I (Coll I), ib onec in (FN) and a mix u e o Coll I and FN (Wine e al., 2011). Figu e 3 shows he s i ening and sp eading o ilamin A-exp essing A7 human melanoma cells and is compa ed wi h di e en s i nesses subs a es coa ed wi h FN o Coll I. S i ening bu no sp eading o A7 melanoma cells depends e y s ongly on whe he in eg ins speci ic o FN o Coll I a e engaged. When A7 cells a e pla ed on gels coa ed wi h sa u a ing amoun s o ei he FN o Coll I, hey sp ead o app oxima ely he same ex en (Figu e 3B), bu he cells on Coll I a e much s i e han hose adhe en o FN (Figu e 3A). When bo h FN and Coll I a e p esen , allowing bo h α1 and β3 in eg ins o bind, adhe en a ea inc eases, bu cell s i ness eaches an in e media e alue be ween hose ound on FN o Coll I alone (Kandow e al., 2007). CELL MIGRATION AND PROGRESSION IN MELANOMA Mo ili y Melanoma s a s wi h he los binding in e cellula connec ions be ween melanocy es and ke a inocy es (Figu e 4). In he p ocess o melanoma de elopmen , due o EMT, cells acqui ed a mesenchymal s em cell pheno ype, con ibu ing o cell mig a ion (Banda chi e al., 2010). The EMT model is e y well explained by Nie o e al. (2016) and B able z e al. (2018), he o ces in ol ed in EMT and i s ela ion wi h cance me as asis a e desc ibed by Wei e al. (2015). A e losing hei connec ions, melanocy es mo e due o a mo ili y p ocess. Mo ili y is he capaci y o a li ing sys em o mo e independen ly, using me abolic ene gy o pe o m mechanical wo k and should no be con used wi h mobili y (Allen, 1981; Figu e 5). Fo cell mig a ion, he e is he con ibu ion o he in e nal o ces (which a e gene a ed by some s uc u es p esen inside each cell, such as he cy oskele al ilamen s) and he ex e nal o ces ( he shea low o blood o lympha ic luids con ibu ions). The mo ili y o cells h ough issues is F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 7Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 8 B ás e al. Melanoma in he Eyes o Mechanobiology FIGURE 3 | Melanoma s i ness (A) and sp eading (B) in unc ion o Collagen ype I (Coll I) and Fib onec in (FN) p esence in hyd ogels. When A7 cells a e pla ed on gels coa ed wi h sa u a ing amoun s o ei he FN o Coll I, hey sp ead o app oxima ely he same ex en (B), bu he cells on Coll I a e much s i e han hose adhe en o FN (A). When bo h FN and Coll I a e p esen , allowing bo h α1 and β3 in eg ins o bind, adhe en a ea inc eases, bu cell s i ness eaches an in e media e alue be ween hose ound on FN o Coll I alone. Each ace d awing inside he cells co esponds o 1 KPa (based on Kandow e al., 2007). a p ocess media ed by in e nal o ces, which a e ela ed wi h he pola iza ion o he cell body wi hin he ECM. This mo ion depends on shape changes, cell-subs a e in e ac ion, and he gene a ion o o ces om he cells media ing h ough he issue. The in e nal o ces esponsible o cell mo ili y a e composed by he ollowing s eps: (i) cy oskele al pola iza ion, o ming ex ensions o ac in ilamen s loca ed in he leading edge; (ii) cell p o usions connec o he ECM sca old by cell su ace adhesion ecep o s; (iii) con ac ion o ac omyosin mo o s ha a e connec ed wi h he ECM subs a es exe ing ac ion o ces; (i ) bipola ension, which is ansmi ed h ough he subs a e, allows he e ac ion o he cell ea comple ing he mo emen o he cell; and ( ) when cells mo e, hey p oduce ECM deg ading p o eases and he ansmig a ed ECM is emodeled by p o eolysis (Tao e al., 2017). Tao e al. (2017) also e e s ha he ac in cy oskele on exhibi s iscous and elas ic esponses, which can be cha ac e ized by he elaxa ion ime obse ed in a c eep esponse a e applying a s ess. The elaxa ion co esponds o he ime scale whe e he cell cy oskele on can low and beha e in an elas ic way. Cy oplasm is mo e han cy oskele on: i has a lo o wa e con en (a ound 70%), which also play a ole in cell mo ion, since wa e lows in be ween he cy oplasm o ganelles. The ex e nal o ces a e ela ed wi h he wa e low and osmo ic p essu e, as well o he luid shea s ess o he blood and lymph. FIGURE 4 | Melanocy es – ke a inocy es adhesion molecules in heal hy and in cance issues. In a heal hy s a e he melanocy e-ke a inocy e in e ac ions a e cha ac e ized by he exis ence o E-cadhe in adhe en junc ions. The ke a inocy es and melanocy es a e connec ed o he ECM h ough he in eg ins. In a cance s a e, he E-cadhe ins a e los and N-cadhe ins a e he adhe en junc ions s ablished be ween melanocy es. Melanocy e dend i es e ac ed and melanocy es acqui ed a ound shape pheno ype. The wa e can pass h ough he cell memb ane ia aquapo in in bo h di ec ions, so he cell can change i s olume, ei he o ced by ex e nal cues like he osmo ic p essu e di e ence be ween inside and ou side o he cell o ac i ely con olled by ion pumps (Tao e al., 2017). Cy oskele on ea angemen occu s h ough he a angemen o h ee p o eins: Rho, Rac and Cdc42. The Rho GTPase amily since hey in luence he appea ance and s uc u e o ilopodia, s ess ibe s and adhesion plaques o melanoma cells (Tao e al., 2017) encou age ac in e ac ion; Rac was ea ed o encou age lamellipodia ex ension bu block ac in depolyme iza ion; Cdc42 was assumed o encou age ilopodia elonga ion and block ac in depolyme iza ion (Tao e al., 2017). Mo ili y is esul ing om wha is calling he mechano ansduc ion “inside-ou ” combining wi h a wa e low in and ou . In human melanoma, by mic opo e il a ion, i was ound ha imen in and ke a in in e media e ilamen exp ession esul s in a mo e mig a o y and in asi e pheno ype (Gao, 2014). The au ho s also ound ha damage o ac in ilamen s and mic o ubules inc eases he de o mabili y con ibu ing o mo ili y and me as iza ion (Gao, 2014). Cell mo ion equi es ha he cy oplasm iscosi y change in he cell pe iphe y h ough he ans o ma ion o he gel s a e o a luidic one. This mechanism also induces o ma ion o p o usions ha F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 8Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 9 B ás e al. Melanoma in he Eyes o Mechanobiology FIGURE 5 | Cell mo ili y: S age 0 (S eady s a e) -Cells main ain he connec ions o he ECM h ough he in eg ins; S age 1 (P o usion o he leading edge) -in e nal o ces induce he F-ac in polyme iza ion and he wa e low en e s h ough he cell memb ane; he ac in polyme iza ion induce he o ma ion o p o usions; S age 2 (Adhesion o leading edge) - he co ex unde ension, accumula es new ac in, h ough he mo emen o unpolyme ized ac in o he p o usion edge, and new adhesion molecules appea in he p o usion pa . S age 3 (Mo emen o he cell body) - he e is he con ac ion o he cell in he same di ec ion o he cell mo emen , and he wa e low exi s om he cell in he opposi e di ec ion, con ibu ing o cell mo ili y; S age 4 (Deadhesion o he ailing edge) - Deadhesion on he cell back edge: he in eg ins in he back o he cell dec eases, and new adhesion molecules appea ed in he on o he cell in he same di ec ion o cell mo emen based on Tao e al. (2017). acili a e cell locomo ion. Ac in ilamen s a e bond o he plasma memb ane by a glycop o ein. In he p esence o his glycop o ein cells a e s able, he p o usions a e well o ganized, occu ing cell mo emen . I his glycop o ein is no p esen , as in he case o ce ain human melanoma, locomo ion is impai ed, and he plasma memb ane shows blebbing (because o he cy oskele on ins abili y). I he p esence o he glycop o ein is e-es ablished, blebbing disappea s and cell mo emen is eco e ed. This glycop o ein also inc eases he s i ness o no mal cells, meaning ha in melanoma cance he lack o his p o ein inc eases he de o mabili y and con ibu es o he e iciency o he mig a ion p ocess o umo cells (Gao, 2014). The whole cell, including he nucleus, mus p esen a ce ain plas ici y. Inside he nucleus, he e is ch oma in oge he wi h lamins in coo dina ion wi h he cy oskele on, main aining shape and mechanical s abili y o he nucleus. Ch oma in wo ks like an elas ic sp ing and is esponsible o he o ce esponse o small de o ma ions o he nucleus. Lamin A de o ms easily o small ex ensions and gi es s i ness o esis la ge nuclea de o ma ions (S ephens e al., 2019). Mic o ubules in he cell cy oskele on exe o ces gene a ed by dynein ha can de o m o e en cause he up u e o he nucleus. Vimen in p o ec s he nucleus gi ing i s abili y and pe inuclea s i ness, which hinde s 3D mo ili y. Ac in also s abilizes he nucleus shape oge he wi h mic o ubules (S ephens e al., 2019). Al e a ions F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 9Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 16 B ás e al. Melanoma in he Eyes o Mechanobiology REFERENCES Ahmed, F., and Haass, N. K. (2018). Mic oen i onmen -d i en dynamic he e ogenei y and pheno ypic plas ici y as a mechanism o melanoma he apy esis ance. F on . Oncol. 8:173. doi: 10.3389/ onc.2018.00173 Allen, R. D. (1981). Mo ili y. J. Cell Biol. 91, 148s–155s. doi: 10.1083/jcb.91.3.148s Anchan, A., Kalogi ou-Baldwin, P., Johnson, R., Kho, D. T., Joseph, W., Hucklesby, J., e al. (2019). Real- ime measu emen o melanoma cell-media ed human b ain endo helial ba ie dis up ion using elec ic cell-subs a e impedance sensing echnology. Biosenso s 9:56. doi: 10.3390/bios9020056 A Melanoma Founda ion (2014). Unde s anding Melanoma. h ps://www. aima melanoma.o g/abou -melanoma/melanoma-de elops/cells- umo s/ (accessed July 14, 2019). Aya-Bonilla, C. A., Ma sa ela, G., F eeman, J. B., Lomma, C., F ank, M. H., Kha ak, M. A., e al. (2017). Isola ion and de ec ion o ci cula ing umou cells om me as a ic melanoma pa ien s using a slan ed spi al mic o luidic de ice. Onco a ge . 8, 67355–67368. doi: 10.18632/onco a ge .18641 Banda chi, B., Jabba i, C. A., Vedadi, A., and Na ab, R. (2013). Molecula biology o no mal melanocy es and melanoma cells. J. Clin. Pa hol. 66, 644–648. doi: 10.1136/jclinpa h-2013-201471 Banda chi, B., Ma, L., Na ab, R., Se h, A., and Ras y, G. (2010). F om melanocy e o me as a ic malignan melanoma. De ma ol. Res. P ac . 2010:583748. doi: 10.1155/2010/583748 Ba , J., Kuma , V., Ro h, W., Schwa z, N., Rich e , M., Leube, R. E., e al. (2014). Skin agili y and impai ed desmosomal adhesion in mice lacking all ke a ins. J. In es ig., De ma ol. 134, 1012–1022. doi: 10.1038/jid.2013.416 Bax e , L. T., and Jain, R. K. (1989). T anspo o luid and mac omolecules in umo s. I. Role o in e s i ial p essu e and con ec ion. Mic o asc. Res. 37, 77–104. doi: 10.1016/0026-2862(89)90074-5 Benne , D., Viswana h, B., Kim, S., and An, J. H. (2017). An ul a-sensi i e biophysical isk assessmen o ligh e ec on skin cells. Onco a ge 8, 47861– 47875. doi: 10.18632/onco a ge .18136 Bikle, D. D. (2004). Vi amin D egula ed ke a inocy e di e en ia ion. J. Cell. Biochem. 92, 436–444. doi: 10.1002/jcb.20095 Bob owska, J., Awsiuk, K., Pabijan, J., Bob owski, P., Lekki, J., Sowa, K. M., e al. (2019). Biophysical and biochemical cha ac e is ics as complemen a y indica o s o melanoma p og ession. Anal. Chem. 91, 9885–9892. doi: 10.1021/ acs.analchem.9b01542 B able z, T., Kallu i, R., Nie o, M. A., and Weinbe g, R. A. (2018). EMT in cance . Na . Re . Cance . 18, 128–134. doi: 10.1038/n c.2017.118 B ad o d, P. T. (2009). Skin cance in skin o colo . De ma ol. Nu s. 21, 170–178. B aeue , R. R., Wa son, I. R., Wu, C. J., Mobley, A. K., Kamiya, T., Shoshan, E., e al. (2013). Why is melanoma so me as a ic? Pigmen Cell Melanoma Res. 27, 19–36. doi: 10.1111/pcm .12172 B ode s-Bondon, F., Nguyen Ho-Bouldoi es, T. H., Fe nandez-Sanchez, M. E., and Fa ge, E. (2018). Mechano ansduc ion in umo p og ession: he da k side o he o ce. J. Cell Biol. 217, 1571–1587. doi: 10.1083/jcb.201701039 Cance Resea ch Ins i u e. (2019). Wha is immuno he apy? A aliable a : h ps: //www.cance esea ch.o g/immuno he apy/wha -is-immuno he apy (accessed July 14, 2019). Checa, S., Rausch, M. K., Pe e sen, A., Kuhl, E., and Duda, G. N. (2015). The eme gence o ex acellula ma ix mechanics and cell ac ion o ces as impo an egula o s o cellula sel -o ganiza ion. Biomech. Model. Mechanobiol.14, 1–13. doi: 10.1007/s10237-014-0581-9 Chelbe g, M. K., Tsiliba y, E. C., Hause , A. R., and Mcca hy, J. B. (1989). Type IV collagen-media ed melanoma cell adhesion and mig a ion: in ol emen o mul iple, dis inc domains o he collagen molecule. Cance Res. 49, 4796. Chen, Y., Suma dika, I. W., Tomonobu, N., Wina sa Ruma, I. M., Kinoshi a, R., Kondo, E., e al. (2019). Melanoma cell adhesion molecule is he d i ing o ce behind he dissemina ion o melanoma upon S100A8/A9 binding in he o iginal skin lesion. Cance Le . 452, 178–190. doi: 10.1016/j.canle .2019.03.023 Chi ukula, V. K., K og, B. L., Nausee , J. T., Hen y, M. D., and Vigmos ad, S. C. (2015). Al e a ions in cance cell mechanical p ope ies a e luid shea s ess exposu e: a mic opipe e aspi a ion s udy. Cell Heal h Cy oskele .7, 25–35. doi: 10.2147/chc.S71852 Choi, H., Kim, M., Ahn, S. I., Cho, E. G., Lee, T. R., and Shin, J. H. (2014). Regula ion o pigmen a ion by subs a e elas ici y in no mal human melanocy es and melano ic MNT1 human melanoma cells. Exp. De ma ol. 23, 172–177. doi: 10.1111/exd.12343 Chung, H., Suh, E.-K., Han, I.-O., and Oh, E.-S. (2011). Ke a inocy e-de i ed laminin-332 p omo es adhesion and mig a ion in melanocy es and melanoma. J. Biol. Chemi. 286, 13438–13447. doi: 10.1074/jbc.M110.166751 Damsky, W. E., Rosenbaum, L. E., and Bosenbe g, M. (2010). Decoding melanoma me as asis. Cance s 3, 126–163. doi: 10.3390/cance s3010126 Daza, R., González-Be múdez, B., C uces, J., De La Fuen e, M., Plaza, G. R., A oyo-He nández, M., e al. (2019). Compa ison o cell mechanical measu emen s p o ided by a omic o ce mic oscopy (AFM) and mic opipe e aspi a ion (MPA). J. Mech. Beha . Biomed. Ma e .95, 103–115. doi: 10.1016/j. jmbbm.2019.03.031 Desja dins-Pa k, H. E., Fos e , D. S., and Longake , M. T. (2018). Fib oblas s and wound healing: an upda e. Regen. Med. 13, 491–495. doi: 10.2217/ me-2018- 0073 Dissanayake, S. K., Wade, M., Johnson, C. E., O’connell, M. P., Leo lela, P. D., F ench, A. D., e al. (2007). The Wn 5A/p o ein kinase C pa hway media es mo ili y in melanoma cells ia he inhibi ion o me as asis supp esso s and ini ia ion o an epi helial o mesenchymal ansi ion. J., Biol. Chemi. 282, 17259–17271. doi: 10.1074/jbc.M700075200 Fa ies, M. B. (2016). in alesional immuno he apy o me as a ic melanoma: he oldes and newes ea men in oncology. C i . Re . Oncog. 21, 65–73. doi: 10.1615/C i Re Oncog.2016017124 Fazakas, C., Wilhelm, I., Nagyoszi, P., Fa kas, A. E., Haskó, J., Molná , J., e al. (2011). T ansmig a ion o melanoma cells h ough he blood-b ain ba ie : ole o endo helial igh junc ions and melanoma- eleased se ine p o eases. PloS One 6:e20758. doi: 10.1371/jou nal.pone.0020758 Feue e , L., Lamm, S., Henz, I., Kappelmann-Fenzl, M., Ha e kamp, S., Meie johann, S., e al. (2019). Role o melanoma inhibi o y ac i i y in melanocy e senescence. Pigmen Cell Melanoma Res. 32, 777–791. doi: 10.1111/ pcm .12801 F anklin, C., Li ings one, E., Roesch, A., Schilling, B., and Schadendo , D. (2017). Immuno he apy in melanoma: Recen ad ances and u u e di ec ions. Eu . J. Su gical Oncol. 43, 604–611. doi: 10.1016/j.ejso.2016.07.145 Gao, H. (2014). P obing mechanical p inciples o cell-nanoma e ial in e ac ions. J. Mech. Phys. Solids 62, 312–339. doi: 10.1016/j.jmps.2013.08.018 Gao, J., Aksoy, B. A., Dog usoz, U., D esdne , G., G oss, B., Sume , S. O., e al. (2013). In eg a i e analysis o complex cance genomics and clinical p o iles using he cBioPo al. Sci. Signal. 6:l1. doi: 10.1126/scisignal.2004088 Go ode sky, R., Sheskin, J., and Wein eb, A. (1986). I on, coppe , and zinc concen a ions in no mal skin and in a ious nonmalignan and malignan lesions. In . J. De ma ol. 25, 440–445. doi: 10.1111/j.1365-4362.1986. b03449.x Haase, K., Macadangdang, J. K., Ed ing on, C. H., Cue ie , C. M., Hadjian oniou, S., Ha den, J. L., e al. (2016). Ex acellula o ces cause he nucleus o de o m in a highly con olled aniso opic manne . Sci. Rep. 6:21300. doi: 10.1038/ s ep21300 Haass, N. K., Smalley, K. S., Li, L., and He lyn, M. (2005). Adhesion, mig a ion and communica ion in melanocy es and melanoma. Pigmen Cell Res. 18, 150–159. doi: 10.1111/j.1600-0749.2005.00235.x Hall, C. J., Boyle, R. H., Sun, X., Wicke , S. M., Misa, J. P., K issansen, G. W., e al. (2014). Epide mal cells help coo dina e leukocy e mig a ion du ing in lamma ion h ough a y acid- uelled ma ix me allop o einase p oduc ion. Na . Commun. 5:3880. doi: 10.1038/ncomms4880 Ha z eld, M., Keil, R., and Magin, T. M. (2017). Desmosomes and in e media e ilamen s: hei consequences o issue mechanics. Cold Sp ing Ha bo Pe spec . Biol. 9:a029157. doi: 10.1101/cshpe spec .a029157 Hege eld , Y., Tusch, M., B ocke , E. B., and F iedl, P. (2002). Collec i e cell mo emen in p ima y melanoma explan s: plas ici y o cell-cell in e ac ion, be a1-in eg in unc ion, and mig a ion s a egies. Cance Res. 62, 2125–2130. He man, H., Fazakas, C., Haskó, J., Molná , K., Mészá os, Á, Nyúl-Tó h, Á, e al. (2019). Pa acellula and anscellula mig a ion o me as a ic cells h ough he ce eb al endo helium. J. Cell. Mol. Med. 23, 2619–2631. doi: 10.1111/jcmm. 14156 He zbe g, B., and Fishe , D. E. (2016). Me as a ic melanoma and immuno he apy. Clin. Immunol. 172, 105–110. doi: 10.1016/j.clim.2016.07.006 Hi obe, T. (2014). Ke a inocy es egula e he unc ion o melanocy es. De ma ol. Sinica 32, 200–204. doi: 10.1016/j.dsi.2014.05.002 F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 16 Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 17 B ás e al. Melanoma in he Eyes o Mechanobiology Hj, X., Zhang, H , Su, K., Yang, J., Wang, H., e al. (2017). E ec o daca bazine on CD44 in li e melanoma cells as measu ed by a omic o ce mic oscopy-based nanoscopy. In . J. Nanomedicine 12, 8867–8886. doi: 10.2147/ijn.s149107 Ho sch oe , V., Koch, K., Ludwig, F., F iedl, P., Obe lei hne , H., S ock, C., e al. (2017). Ex acellula p o ona ion modula es cell-cell in e ac ion mechanics and issue in asion in human melanoma cells. Sci. Rep. 7:42369. doi: 10.1038/ s ep42369 Hsu, C. K., Lin, H. H., Ha n, H. I., Hughes, M. W., Tang, M. J., and Yang, C. C. (2018). Mechanical o ces in skin diso de s. J. De ma ol. Sci. 90, 232–240. doi: 10.1016/j.jde msci.2018.03.004 Huang, J., Wu, S., Ba e a, J., Ma hews, K., and Pan, D. (2005). The hippo signaling pa hway coo dina ely egula es cell p oli e a ion and apop osis by inac i a ing Yo kie, he D osophila Homolog o YAP. Cell 122, 421–434. doi: 10.1016/j.cell. 2005.06.007 Huang, Q., Hu, X., He, W., Zhao, Y., Hao, S., Wu, Q., e al. (2018). Fluid shea s ess and umo me as asis. Am. J. Cance Res. 8, 763–777. I o, Y., Kanama u, A., and Tada, A. (2006). Cen au eidin p omo es dend i e e ac ion o melanocy es by ac i a ing Rho. Bioch. Biophys. Ac a 1760, 487– 494. doi: 10.1016/j.bbagen.2006.01.003 Jackson, B., B own, S. J., A ilion, A. A., O’Shaughnessy, R. F., Sully, K., and Akindu o, O. (2011). ) TALE homeodomain p o eins egula e si e-speci ic e minal di e en ia ion, LCE genes and epide mal ba ie . J. Cell Sci. 124, 1681–1690. doi: 10.1242/jcs.077552 Jain, R. K. (1988). De e minan s o umo blood low: a e iew. Cance Res. 48, 2641–2658. Jain, R. K., and Bax e , L. T. (1988). Mechanisms o he e ogeneous dis ibu ion o monoclonal an ibodies and o he mac omolecules in umo s: signi icance o ele a ed in e s i ial p essu e. Cance Res. 48, 7022–7032. Jain, R. K., Ma in, J. D., and S ylianopoulos, T. (2014). The ole o mechanical o ces in umo g ow h and he apy. Annu. Re i. Biomed. Eng. 16, 321–346. doi: 10.1146/annu e -bioeng-071813-105259 Johnson, D. B., Smalley, K. S., and Sosman, J. A. (2014). Molecula pa hways: a ge ing NRAS in melanoma and acu e myelogenous leukemia. Clin. Cance Res. 20, 4186–4192. doi: 10.1158/1078-0432.Cc -13-3270 Ju, R. J., S ehbens, S. J., and Haass, N. K. (2018). The Role o Melanoma Cell- S oma In e ac ion in Cell Mo ili y, In asion, and Me as asis. F on . Med. 5:307. doi: 10.3389/ med.2018.00307 Kandow, C. E., Geo ges, P. C., Janmey, P. A., and Beningo, K. A. (2007). Polyac ylamide hyd ogels o cell mechanics: s eps owa d op imiza ion and al e na i e uses. Me hods Cell Biol.83, 29–46. doi: 10.1016/s0091-679x(07) 83002-0 Ka an za, V. (2011). Ke a ins in heal h and cance : mo e han me e epi helial cell ma ke s. Oncogene 30, 127–138. doi: 10.1038/onc.2010.456 Kasas, S. L., and Die le , G. (2013). Mechanical p ope ies o biological specimens explo ed by a omic o ce mic oscopy. J. Phys. D: Appl. Phys. 46:133001. doi: 10.1088/0022-3727/46/13/133001 Kawakubo, T., Yasukochi, A., Okamo o, K., Okamo o, Y., Nakamu a, S., and Yamamo o, K. (2011). The ole o ca hepsin E in e minal di e en ia ion o ke a inocy es. Biol. Chem. 392, 571–585. doi: 10.1515/BC.2011.060 Kim, B. J., Lee, S. Y., Jee, S., A ajano , A., and Yang, S. (2017). Mic o- iscome e o measu ing shea - a ying blood iscosi y o e a wide- anging shea a e. Senso s 17:1442. doi: 10.3390/s17061442 Kim, T. H., Rowa , A. C., and Sloan, E. K. (2016). Neu al egula ion o cance : om mechanobiology o in lamma ion. Clin. T ansl. Immunol. 5:e78. doi: 10. 1038/c i.2016.18 Kippenbe ge , S., Be nd, A., Be ei e -Hahn, J., Rami ez-Bosca, A., and Kau mann, R. (1998). The mechanism o melanocy e dend i e o ma ion: he impac o di e en ia ing ke a inocy es. Pigmen Cell Res. 11, 34–37. doi: 10.1111/j.1600- 0749.1998. b00708.x Klemke, M., Weschen elde , T., Kons andin, M. H., and Sams ag, Y. (2007). High a ini y in e ac ion o in eg in alpha4be a1 (VLA-4) and ascula cell adhesion molecule 1 (VCAM-1) enhances mig a ion o human melanoma cells ac oss ac i a ed endo helial cell laye s. J. Cell Physiol. 212, 368–374. doi: 10.1002/jcp. 21029 Kode , O. D. B., Bendlo a, B., Syko o a, V., K ajso a, I., S o k, J., Kuce a, J., e al. (2018). Mic oen i onmen d i en esis ance o BRa inhibi ion in a melanoma pa ien is accompanied by b oad changes o gene me hyla ion and exp ession in dis al ib oblas s. In . J. Mol. Med. 41, 2687–2703. doi: 10.3892/ijmm.2018. 3448 Kolasi´ nska, E., Janik, M. E., Li yñska, A., and P zybyło, M. (2019). Con ibu ion o sialic acids o in eg in α5β1 unc ioning in melanoma cells. Ad . Med Sci. 64, 267–273. doi: 10.1016/j.ad ms.2019.02.002 Koumou sakos, P., Pi kin, I., and Milde, F. (2013). The luid mechanics o cance and i s he apy. Annu. Re . Fluid Mech. 45:30. doi: 10.1146/annu e - luid- 120710-101102 Koza , I., Ma gue, C., Ro henga e , S., Haan, C., and K eis, S. (2019). Many ways o esis ance: how melanoma cells e ade a ge ed he apies. Biochim. Biophys. Ac a (BBA) Re . Cance 1871, 313–322. doi: 10.1016/j.bbcan.2019.02.002 K use, P. F. J ., and Pa e son, M. K. (1973). Tissue Cul u e Me hods and Applica ions. New Yo k: Academic P ess. Kunz, M. (2014). Oncogenes in melanoma: an upda e. Eu . J. Cell Biol. 93, 1–10. doi: 10.1016/j.ejcb.2013.12.002 Kwa, M., He um, K., and B akebusch, C. (2019). Cance -associa ed ib oblas s: how do hey con ibu e o me as asis? Clin. Exp. Me as asis 36, 71–86. doi: 10.1007/s10585-019-09959-0 Lassalle, M. W., Iga ashi, S., Sasaki, M., Wakama su, K., I o, S., and Ho ikoshi, T. (2003). E ec s o melanogenesis-inducing ni ic oxide and his amine on he p oduc ion o eumelanin and pheomelanin in cul u ed human melanocy es. Pigmen Cell Res. 16, 81–84. doi: 10.1034/j.1600-0749.2003.00004.x Lazo a, R., and Pawelek, J. M. (2009). Why do melanomas ge so da k? Exp. De ma ol. 18, 934–938. doi: 10.1111/j.1600-0625.2009.00933.x Lee, G., Han, S.-B., Lee, J.-H., Kim, H.-W., and Kim, D.-H. (2019). Cance mechanobiology: mic oen i onmen al sensing and me as asis. ACS Bioma e . Sci. Eng. 5, 3735–3752. doi: 10.1021/acsbioma e ials.8b01230 Lee, H. J., Diaz, M. F., P ice, K. M., Ozuna, J. A., Zhang, S., Se ick-Mu aca, E. M., e al. (2017). Fluid shea s ess ac i a es YAP1 o p omo e cance cell mo ili y. Na . Commun. 8, 14122. doi: 10.1038/ncomms14122 Lekka, M., Gil, D., Pogoda, K., Duliñska-Li ewka, J., Jach, R., Gos ek, J., e al. (2012). Cance cell de ec ion in issue sec ions using AFM. A ch. Biochem. Biophys. 518, 151–156. doi: 10.1016/j.abb.2011.12.013 Li, G., Sa yamoo hy, K., Meie , F., Be king, C., Bogen iede , T., and He lyn, M. (2003). Func ion and egula ion o melanoma-s omal ib oblas in e ac ions: when seeds mee soil. Oncogene 22, 3162–3171. doi: 10.1038/sj.onc.1206455 Li, X., Regezi, J., Ross, F. P., Blys one, S., Ilic, D., Leong, S. P., e al. (2001). In eg in alpha be a3 media es K1735 mu ine melanoma cell mo ili y in i o and in i o. J. Cell Sci. 114, 2665–2672. Makale, M. (2007). Cellula mechanobiology and cance me as asis. Bi h De ec s Res. Pa C Emb yo Today 81, 329–343. doi: 10.1002/bd c.20110 Mangahas, C. R., Dela C uz, G. V., Schneide , R. J., and Jamal, S. (2004). Endo helin-1 up egula es MCAM in melanocy es. J. In es ig. De ma ol. 123, 1135–1139. doi: 10.1111/j.0022-202X.2004.23480.x Ma sa ela, G., Aya-Bonilla, C. A., Wa kiani, M. E., G ay, E. S., and Ziman, M. (2018). Melanoma ci cula ing umo cells: bene i s and challenges equi ed o clinical applica ion. Cance Le . 424, 1–8. doi: 10.1016/j.canle .2018.03.013 McG a h, J. A., Eady, R. A. J., and Pope, F. M. (2004). Ana omy and O ganiza ion o Human Skin. Blackwell Publishing. Mi, K. (2009). “Making an epide mis,” in In e na ional Symposium on Ol ac ion and Tas e, ed. T. E. Finge (New Yo k, NY: Academy o Sciences), 4. Mocellin, S., Keilholz, U., Rossi, C. R., and Ni i, D. (2006). Ci cula ing umo cells: he ‘leukemic phase’ o solid cance s. T ends Mol. Medi. 12, 130–139. doi: 10.1016/j.molmed.2006.01.006 Mo iceau, G., Hugo, W., Hong, A., Shi, H., Kong, X., Yu, C. C., e al. (2015). Tunable-combina o ial mechanisms o acqui ed esis ance limi he e icacy o BRAF/mek co a ge ing bu esul in melanoma d ug addic ion. Cance Cell. 27, 240–256. doi: 10.1016/j.ccell.2014.11.018 Mum o d, B. S., and Robe son, G. P. (2014). Ci cula ing melanoma cells in he diagnosis and moni o ing o melanoma: an app aisal o clinical po en ial. Mol Diagn The . 18, 175–183. doi: 10.1007/s40291-013-0071-2 Nie o, M. A., Huang, R. Y., Jackson, R. A., and Thie y, J. P. (2016). EMT: 2016. Cell 166, 21–45. doi: 10.1016/j.cell.2016.06.028 Nikolo ska, K., Spillmann, D., Haie , J., Ladányi, A., S ock, C., and Seidle , D. G. (2017). Melanoma cell adhesion and mig a ion is modula ed by he u onyl 2-O sul o ans e ase. PloS One 12:e0170054. doi: 10.1371/jou nal.pone.0170054 O ioli, D., and Dellamb a, E. (2018). Epigene ic egula ion o skin cells in na u al aging and p ema u e aging diseases. Cells 7, E268. doi: 10.3390/cells7120268 Pade a, T.P., S oll, B. R., Too edman, J. B., Capen, D., Di Tomaso, E., and Jain, R. K. (2004). Pa hology: cance cells comp ess in a umou essels. Na u e 427:695. doi: 10.1038/427695a F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 17 Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 18 B ás e al. Melanoma in he Eyes o Mechanobiology Paluncic, J., Ko ace ic, Z., Jansson, P. J., Kalinowski, D., Me lo , A. M., Huang, M. L., e al. (2016). Roads o melanoma: Key pa hways and eme ging playe s in melanoma p og ession and oncogenic signaling. Biochim. Biophys. Ac a Mol. Cell Res. 1863, 770–784. doi: 10.1016/j.bbamc .2016.01.025 Pe i cle c, E., S omblad, S., Von Schalscha, T. L., Mi jans, F., Piula s, J., Mon gome y, A. M., e al. (1999). In eg in alpha( )be a3 p omo es M21 melanoma g ow h in human skin by egula ing umo cell su i al. Cance Res. 59, 2724–2730. Pie as, K., and Os man, A. (2010). Hallma ks o cance : in e ac ions wi h he umo s oma. Exp Cell Res. 316, 1324–1331. doi: 10.1016/j.yexc .2010.02.045 P oksch, E., B andne , J. M., and Jensen, J. M. (2008). The skin: an indispensable ba ie. Exp. De ma ol. 17, 1063–1072. doi: 10.1111/j.1600-0625.2008.00786.x Pugh, T. J., Mo ozo a, O., A iyeh, E. F., Asgha zadeh, S., Wei, J. S., and Auclai , D. (2013). The gene ic landscape o high- isk neu oblas oma. Na . Gene . 45, 279–284. doi: 10.1038/ng.2529 Qi, J., Chen, N., Wang, J., and Siu, C.-H. (2005). T ansendo helial mig a ion o melanoma cells in ol es N-cadhe in-media ed adhesion and ac i a ion o he be a-ca enin signaling pa hway. Mol. Biol. Cell 16, 4386–4397. doi: 10.1091/ mbc.e05-03-0186 Qiu, Y., Mye s, D. R., and Lam, W. A. (2019). The biophysics and mechanics o blood om a ma e ials pe spec i e. Na . Re . Ma e . 294–311. doi: 10.1038/ s41578-019-0099-y Ramms, L., Fab is, G., Windo e , R., Schwa z, N., Sp inge , R., Zhou, C., e al. (2013). Ke a ins as he main componen o he mechanical in eg i y o ke a inocy es. P oc. Na l. Acad. Sci. U.S.A. 110, 18513–18518. doi: 10.1073/pnas. 1313491110 Rebhun, R. B., Cheng, H., Ge shenwald, J. E., Fan, D., Fidle , I. J., and Langley, R. R. (2010). Cons i u i e exp ession o he alpha4 in eg in co ela es wi h umo igenici y and lymph node me as asis o he B16 mu ine melanoma. Neoplasia 12, 173–182. doi: 10.1593/neo.91604 Rheinwald, J. G., and G een, H. (1975). Se ial cul i a ion o s ains o human epide mal ke a inocy es: he o ma ion o ke a inizing colonies om single cells. Cell 6, 331–343. doi: 10.1016/S0092-8674(75)80001-8 Roca-Cusachs, P., Gau hie , N. C., Del Rio, A., and Shee z, M. P. (2009). Clus e ing o alpha(5)be a(1) in eg ins de e mines adhesion s eng h whe eas alpha( )be a(3) and alin enable mechano ansduc ion. P oc. Na l. Acade. Sci. U.S.A. 106, 16245–16250. doi: 10.1073/pnas.0902818106 Rod iguez-Ce dei a, C., Ca ne o G ego io, M., Lopez-Ba cenas, A., Sanchez- Blanco, E., Sanchez-Blanco, B., Fabb ocini, G., e al. (2017). Ad ances in immuno he apy o melanoma: a comp ehensi e e iew. Med. In lamm. 2017, 3264217. doi: 10.1155/2017/3264217 Sanlo enzo, M., Vujic, I., Posch, C., Dajee, A., Yen, A., and Kim, S. (2014). Melan- oma immuno he apy. Cance Biol. The .15, 665–674. doi: 10.4161/cb .28555 Sa na, M., Zadlo, A., He manowicz, P., Madeja, Z., Bu da, K., and Sa na, T. (2014). Cell elas ici y is an impo an indica o o he me as a ic pheno ype o melanoma cells. Exp. De ma ol. 23, 813–818. doi: 10.1111/exd.12535 Sa na, M., Zadlo, A., Pila , A., Olchawa, M., Gkogkolou, P., Bu da, K., e al. (2013). Nanomechanical analysis o pigmen ed human melanoma cells. Pigmen Cell Melanoma Res. 26, 727–730. doi: 10.1111/pcm .12113 Sco , G. (2002). Rac and ho: he s o y behind melanocy e dend i e o ma ion. Pigmen Cell Res. 15, 322–330. doi: 10.1034/j.1600-0749.2002.02056.x Shin, M., Lim, H., Kang, B., Lee, G., Uhm, Y., and Lee, M. (2014). Nanos uc u al change o human melanocy es s imula ed wi h alpha-melanocy e s imula ing ho mone obse ed by a omic o ce mic oscopy. Scanning 36, 570–575. doi: 10.1002/sca.21156 Sil e , F., and Sipe ko, L. (2003). Mechanosensing and mechanochemical ansduc ion: how is mechanical ene gy sensed and con e ed in o chemical ene gy in an ex acellula ma ix? C i . Re . Biomed Eng. 31, 255–331. doi: 10.1615/C i Re BiomedEng. 31.i4.10 Slominski, R. M., Zmijewski, M. A., and Slominski, A. T. (2015). The ole o melanin pigmen in melanoma. Exp. De ma ol. 24, 258–259. doi: 10.1111/exd. 12618 S a k, M. S., Bonazzi, V. F., Boyle, G. M., Palme , J. M., Symmons, J., Lanagan, C. M., e al. (2015). miR-514a egula es he umou supp esso NF1 and modula es BRAFi sensi i i y in melanoma. Onco a ge 6, 17753–17763. doi: 10.18632/onco a ge .3924 S ephens, A. D., Banigan, E. J., and Ma ko, J. F. (2019). Ch oma in’s physical p ope ies shape he nucleus and i s unc ions. Cu . Opi. Cell Biol. 58, 76–84. doi: 10.1016/j.ceb.2019.02.006 S aussman, R., Mo ikawa, T., Shee, K., Ba zily-Rokni, M., Qian, Z. R., Du, J., e al. (2012). Tumou mic o-en i onmen elici s inna e esis ance o RAF inhibi o s h ough HGF sec e ion. Na u e 487, 500–504. doi: 10.1038/na u e 11183 S ylianopoulos, T., and Jain, R. K. (2013). Combining wo s a egies o imp o e pe usion and d ug deli e y in solid umo s. P oc. Na l. Acad. Sci. U. S. A. 110, 18632–18637. doi: 10.1073/pnas.1318415110 Swamina han, V., My h eye, K., Tim O’b ien, E., Be chuck, A., Blobe, G. C., and Supe ine, R. (2011). Mechanical S i ness g ades me as a ic po en ial in pa ien umo cells and in cance cell lines. Cance Res. 71, 5075–5080. doi: 10.1158/ 0008-5472.CAN-11-0247 Tan, Y., Kong, C. W., Chen, S., Cheng, S. H., Li, R. A., and Sun, D. (2012). P obing he mechanobiological p ope ies o human emb yonic s em cells in ca diac di e en ia ion by op ical weeze s. J. Biomech. 45, 123–128. doi: 10. 1016/j.jbiomech.2011.09.007 Tanne , K. (2018). Pe spec i e: The ole o mechanobiology in he e iology o b ain me as asis. APL Bioeng 2, 031801. doi: 10.1063/1.5024394 Tao, J., Li, Y., Vig, D. K., and Sun, S. X. (2017). Cell mechanics: a dialogue. Rep. P og. Phys. 80:036601. Tawbi, H., Gli za Oli a, I. C., and Sch a sman, G. (2018). Ad ances in he sys emic ea men o melanoma b ain me as ases. Anna. Oncol. 29, 1509–1520. doi: 10.1093/annonc/mdy185 Tue enbe g, A., Schmi , E., Knop, J., and Jonulei , H. (2007). Dend i ic cell-based immuno he apy o malignan melanoma: success and limi a ions. J. Deu sch. De ma ol. Ges. 5, 190–196. doi: 10.1111/j.1610-0387.2007.06179.x Uong, A., and Zon, L. I. (2010). Melanocy es in de elopmen and cance . J. Cell Physiol. 222, 38–41. doi: 10.1002/jcp.21935 Van Den Bossche, K., Naeyae , J. M., and Lambe , J. (2006). The ques o he mechanism o melanin ans e . T a ic 7, 769–778. doi: 10.1111/j.1600-0854. 2006.00425.x Vielmu h, F., Wanuske, M. T., Rade a, M. Y., Hie maie , M., Kugelmann, D., Wal e , E., e al. (2018). Ke a ins egula e he adhesi e p ope ies o desmosomal cadhe ins h ough signaling. J. In es ig. De ma ol. 138, 121–131. doi: 10.1016/j. jid.2017.08.033 Viji Babu, P. K., Rianna, C., Belge, G., Mi as schijski, U., and Radmache , M. (2018). Mechanical and mig a o y p ope ies o no mal, sca , and dupuy en’s ib oblas s. J. Mol. Recogn 31:e2719. doi: 10.1002/jm .2719 Wang, A. S., and D eesen, O. (2018). Bioma ke s o cellula senescence and skin aging. F on . Gene . 9:247. doi: 10.3389/ gene.2018.00247 Wang, J. X., Fukunaga-Kalabis, M., and He lyn, M. (2016). C oss alk in skin: melanocy es, ke a inocy es, s em cells, and melanoma. J. Cell Commun. Signal. 10, 191–196. doi: 10.1007/s12079-016-0349-3 Wang, W. C., Zhang, X. F., Peng, J., Li, X. F., Wang, A. L., Bie, Y. Q., e al. (2018). Su i al mechanisms and in luence ac o s o ci cula ing umo cells. BioMed Res. In . 201:6304701. doi: 10.1155/2018/6304701 Wang, Y., Vienne , C., Robin, S., Be hon, J. Y., He, L., and Humbe , P. (2017). P ecise ole o de mal ib oblas s on melanocy e pigmen a ion. J. De ma ol. Sci. 88, 159–166. doi: 10.1016/j.jde msci.2017.06.018 Wäs e , P., E iksson, I., Vainikka, L., Rosdahl, I., and Öllinge , K. (2016). Ex acellula esicles a e ans e ed om melanocy es o ke a inocy es a e UVA i adia ion. Sci. Rep. 6:27890. doi: 10.1038/s ep27890 Wede , G., Hend iks-Balk, M. C., Smajda, R., Rimoldi, D., Liley, M., Heinzelmann, H., e al. (2014). Inc eased plas ici y o he s i ness o melanoma cells co ela es wi h hei acquisi ion o me as a ic p ope ies. Nanomedicine 10, 141–148. doi: 10.1016/j.nano.2013.07.007 Wei, S. C., Fa e , L., and Yang, J. (2015). The o ces behind EMT and umo me as asis. Cell Cycle14, 2387–2388. doi: 10.1080/15384101.2015.1063296 Wen, S. J., Zhang, W., Ni, N. N., Wu, Q., Wang, X. P., Lin, Y. K., e al. (2017). Exp ession o Rho GTPases amily in melanoma cells and i s in luence on cy oskele on and mig a ion. Onco a ge 8, 30112–30122. doi: 10.18632/ onco a ge .15618 Wine , J. P., Chop a, A., K esh, J. Y., and Janmey, P. A. (2011). “Janmey. Subs a e Elas ici y as a P obe o Measu e Mechanosensing a Cell-Cell and Cell-Ma ix Junc ion,” in Mechanobiology o Cell-Cell and Cell-Ma ix In e ac ions, eds A. W. Johnson and B. A. C. Ha ley (Be lin: Sp inge Science+Business Media). Wu, X. S., Masedunskas, A., Weige , R., Copeland, N. G., Jenkins, N. A., and Hamme , J. A. (2012). Melano egulin egula es a shedding mechanism ha d i es melanosome ans e om melanocy es o ke a inocy es. P oc. Na l. Acad. Sci. U.S.A. 109, E2101–E2109. doi: 10.1073/pnas.1209397109 F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 18 Feb ua y 2020 | Volume 8 | A icle 54 cell-08-00054 Feb ua y 11, 2020 Time: 16:43 # 19 B ás e al. Melanoma in he Eyes o Mechanobiology Yoshioka, S., Fujiwa a, H., Higuchi, T., Yamada, S., Maeda, M., and Fujii, S. (2003). Melanoma cell adhesion molecule (MCAM/CD146) is exp essed on human lu einizing g anulosa cells: enhancemen o i s exp ession by hCG, in e leukin-1 and umou nec osis ac o -alpha. Mol. Hum. Rep od. 9, 311–319. doi: 10.1093/moleh /gag042 Za koob, H., Boddulu i, S., Ponnalu i, S. V., Selby, J. C., and Sande , E. A. (2015). Subs a e s i ness a ec s human ke a inocy e colony o ma ion. Cel. Mol. Bioeng. 8, 32–50. doi: 10.1007/s12195-015-0377-8 Zhang, R.-Z., Zhu, W. Y., Xia, M. Y., and Feng, Y. (2004). Mo phology o cul u ed human epide mal melanocy es obse ed by a omic o ce mic oscopy. Pigmen Cell Res. 17, 62–65. doi: 10.1046/j.1600-0749.2003.00111.x Con lic o In e es : The au ho s decla e ha he esea ch was conduc ed in he absence o any comme cial o inancial ela ionships ha could be cons ued as a po en ial con lic o in e es . Copy igh © 2020 B ás, Radmache , Sousa and G anja. This is an open-access a icle dis ibu ed unde he e ms o he C ea i e Commons A ibu ion License (CC BY). The use, dis ibu ion o ep oduc ion in o he o ums is pe mi ed, p o ided he o iginal au ho (s) and he copy igh owne (s) a e c edi ed and ha he o iginal publica ion in his jou nal is ci ed, in acco dance wi h accep ed academic p ac ice. No use, dis ibu ion o ep oduc ion is pe mi ed which does no comply wi h hese e ms. F on ie s in Cell and De elopmen al Biology | www. on ie sin.o g 19 Feb ua y 2020 | Volume 8 | A icle 54