evaluating-effectiveness-cancer-drug-sensitization-vitro

Evaluating the Effectiveness of Cancer Drug Sensitization <em>In Vitro</em> and <em>In Vivo</em>

Due to the high level of heterogeneity and mutations inherent in human cancers, single agent therapies, or combination regimens which target the same ...

Western University

Gender influences the cardiac response to prolonged increases in workload, with differences at structural, functional, and molecular levels. However, it is unknown if post-ischemic function or metabolism of female hypertrophied hearts differ from male hypertrophied hearts. Thus, we tested the hypothesis that gender influences post-ischemic function of pressure-overload hypertrophied hearts and determined if the effect of gender on post-ischemic outcome could be explained by differences in metabolism, especially the catabolic fate of glucose.

Gender and post-ischemic recovery of hypertrophied rat hearts.

Exercise increases tissue-type plasminogen activator expression in rat cardiomyocytes.

Early breast cancer progression involves advancement through specific morphological stages including atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS) and invasive mammary carcinoma (IMC), although not necessarily always in a linear fashion. Observational studies have examined genetic, epigenetic and gene expression differences in breast tissues representing these stages of progression, but model systems which would allow for experimental testing of specific factors influencing transition through these stages are scarce. The 21T series cell lines, all originally derived from the same patient with metastatic breast cancer, have been proposed to represent a mammary tumor progression series. We report here that three of the 21T cell lines indeed mimic specific stages of human breast cancer progression (21PT-derived cells, ADH; 21NT-derived cells, DCIS; 21MT-1 cells, IMC) when grown in the mammary fat pad of nude mice, albeit after a year. To develop a more rapid, readily manipulatable in vitro assay for examining the biological differences between these cell lines, we have used a 3D Matrigel system. When the three cell lines were grown in 3D Matrigel, they showed characteristic morphologies, in which quantifiable aspects of stage-specific in vivo behaviors (ie, differences in acinar structure formation, cell polarization, colony morphology, cell proliferation, cell invasion) were recapitulated in a reproducible fashion. Gene expression profiling revealed a characteristic pattern for each of the three cell lines. Interestingly, Wnt pathway alterations are particularly predominant in the early transition from 21PTci (ADH) to 21NTci (DCIS), whereas alterations in expression of genes associated with control of cell motility and invasion phenomena are more prominent in the later transition of 21NTci (DCIS) to 21MT-1 (IMC). This system thus reveals potential therapeutic targets and will provide a means of testing the influences of identified genes on transitions between these stages of pre-malignant to malignant growth.

Human 21T breast epithelial cell lines mimic breast cancer progression in vivo and in vitro and show stage-specific gene expression patterns.

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The analysis of dynamic events in the tumor microenvironment during cancer progression is limited by the complexity of current in vivo imaging models. This is coupled with an inability to rapidly modulate and visualize protein activity in real time and to understand the consequence of these perturbations in vivo. We developed an intravital imaging approach that allows the rapid induction and subsequent depletion of target protein levels within human cancer xenografts while assessing the impact on cell behavior and morphology in real time. A conditionally stabilized fluorescent E-cadherin chimera was expressed in metastatic breast cancer cells, and the impact of E-cadherin induction and depletion was visualized using real-time confocal microscopy in a xenograft avian embryo model. We demonstrate the assessment of protein localization, cell morphology and migration in cells undergoing epithelial-mesenchymal and mesenchymal-epithelial transitions in breast tumors. This technique allows for precise control over protein activity in vivo while permitting the temporal analysis of dynamic biophysical parameters.

Imaging the impact of chemically inducible proteins on cellular dynamics in vivo.

L&#x26;#39;analyse des &#xE9;v&#xE9;nements dynamiques dans le microenvironnement de la tumeur lors de la progression du cancer est limit&#xE9; par la complexit&#xE9; du courant dans les mod&#xE8;les d&#x26;#39;imagerie in vivo. Ceci est coupl&#xE9; avec une incapacit&#xE9; &#xE0; moduler rapidement et de visualiser l&#x26;#39;activit&#xE9; des prot&#xE9;ines en temps r&#xE9;el et &#xE0; comprendre les cons&#xE9;quences de ces perturbations in vivo. Nous avons d&#xE9;velopp&#xE9; une approche d&#x26;#39;imagerie intravitale qui permet l&#x26;#39;induction rapide et de l&#x26;#39;&#xE9;puisement apr&#xE8;s des taux de prot&#xE9;ines cibles au sein de x&#xE9;nogreffes de cancer de l&#x26;#39;homme, tout en &#xE9;valuant l&#x26;#39;impact sur le comportement des cellules et la morphologie en temps r&#xE9;el. Une condition stabilis&#xE9;e fluorescente E-cadh&#xE9;rine chim&#xE8;re a &#xE9;t&#xE9; exprim&#xE9;e en cellules canc&#xE9;reuses mammaires m&#xE9;tastatiques, et l&#x26;#39;impact de la E-cadh&#xE9;rine &#xE0; induction et de l&#x26;#39;appauvrissement a &#xE9;t&#xE9; visualis&#xE9;e &#xE0; l&#x26;#39;aide en temps r&#xE9;el microscopie confocale dans un mod&#xE8;le de x&#xE9;nogreffe embryon aviaire. Nous d&#xE9;montrons l&#x26;#39;&#xE9;valuation de la localisation de la prot&#xE9;ine, la morphologie et la migration cellulaire dans les cellules subissant des transitions &#xE9;pith&#xE9;lium-m&#xE9;senchyme et m&#xE9;senchymateuses-&#xE9;pith&#xE9;liale dans les tumeurs du sein. Cette technique permet un contr&#xF4;le pr&#xE9;cis sur l&#x26;#39;activit&#xE9; des prot&#xE9;ines in vivo, tout en permettant l&#x26;#39;analyse temporelle des param&#xE8;tres dynamiques biophysiques.

Imagerie de l&#x26;#39;impact des prot&#xE9;ines inductibles chimiquement sur la dynamique cellulaire in vivo

Die Analyse der dynamischen Ereignisse in den Tumor Mikromilieu w&auml;hrend Fortschreiten wird durch die Komplexit&auml;t der aktuellen in-vivo Bildgebung Modelle begrenzt. Dies ist gekoppelt mit einer Unf&auml;higkeit zu schnell modulieren und Protein-Aktivit&auml;t in Echtzeit zu visualisieren und zu verstehen, die Konsequenz aus diesen Perturbationen in-vivo. Wir entwickelten einen intravital Bildgebung Ansatz, der schnelle Induktion und nachfolgende Ersch&ouml;pfung der Zielwerte Protein im menschlichen Krebs Xenografts w&auml;hrend der Bewertung der Auswirkungen auf das Verhalten der Zelle und Morphologie in Echtzeit erm&ouml;glicht. Bedingt stabilisierte fluoreszierende E-Cadherin Chim&auml;re fand ihren Niederschlag in metastasierendem Brustkrebs-Zellen, und die Auswirkungen von E-Cadherin-Induktion und Ersch&ouml;pfung war mit Echtzeit-konfokale Mikroskopie in einem Xenograft-Vogelgrippe Embryo Modell visualisiert. Wir demonstrieren die Beurteilung der Proteinlokalisation, Zellmorphologie und Migration in Zellen, die &Uuml;bergangsphase zwischen epitheliale-Mesenchymale und Mesenchymale-epitheliale befinden in Brusttumoren. Diese Technik erm&ouml;glicht pr&auml;zise Kontrolle &uuml;ber Protein Aktivit&auml;t in-vivo w&auml;hrend erlaubt die zeitliche Analyse der dynamischen biophysikalische Parameter.

Die Auswirkungen der chemisch durch Induktion Proteine auf Zellul&auml;re Dynamik in-vivo Imaging.

L&#39;analisi di eventi dinamici nel microambiente tumorale durante la progressione del cancro &egrave; limitata dalla complessit&agrave; dei modelli attuali di imaging in vivo. Questo &egrave; accoppiato con un&#39;incapacit&agrave; di rapidamente modulare e visualizzare l&#39;attivit&agrave; della proteina in tempo reale e capire la conseguenza di queste perturbazioni in vivo. Abbiamo sviluppato un approccio intravital imaging che permette la rapida induzione e conseguente deplezione dei livelli di proteina bersaglio all&#39;interno di xenotrapianti tumorali umane durante la valutazione dell&#39;impatto sul comportamento delle cellule e morfologia in tempo reale. Una chimera di E-caderina fluorescente condizionalmente stabilizzata &egrave; stato espresso in cellule di cancro al seno metastatico, e l&#39;impatto dell&#39;induzione E-caderina e deplezione &egrave; stato visualizzato mediante microscopia confocale in tempo reale in un modello di embrione aviaria xenoinnesto. Dimostriamo di localizzazione della proteina, la morfologia cellulare e la migrazione nelle cellule epiteliali-mesenchimali ed epiteliali-mesenchimali transizioni in tumori al seno in fase di valutazione. Questa tecnica permette un controllo preciso sulla attivit&agrave; della proteina in vivo pur consentendo l&#39;analisi temporale dei parametri biofisici dinamici.

L&#39;impatto delle proteine inducibili chimicamente sulle dinamiche cellulare in vivo di Imaging.

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&#xC554; &#xC9C4;&#xD589; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xC885;&#xC591; microenvironment&#xC5D0;&#xC11C; &#xB3D9;&#xC801; &#xC774;&#xBCA4;&#xD2B8;&#xC758; &#xBD84;&#xC11D;&#xC740; &#xD604;&#xC7AC; in vivo &#xC774;&#xBBF8;&#xC9D5; &#xBAA8;&#xB378;&#xC758; &#xBCF5;&#xC7A1;&#xC131;&#xC5D0; &#xC758;&#xD574; &#xC81C;&#xD55C; &#xB429;&#xB2C8;&#xB2E4;. &#xC774;&#xAC83;&#xC740; &#xC2E0;&#xC18D; &#xD558; &#xAC8C; &#xC870;&#xC808; &#xB2E8;&#xBC31;&#xC9C8; &#xD65C;&#xB3D9; &#xC2E4;&#xC2DC;&#xAC04;&#xC73C;&#xB85C; &#xC2DC;&#xAC01;&#xD654; &#xD558; &#xACE0; &#xC774;&#xB7EC;&#xD55C; &#xBB3C;&#xACB0; vivo&#xC5D0;&#xC11C;&#xC758; &#xACB0;&#xACFC; &#xC774;&#xD574; &#xD558;&#xB294; &#xBB34; &#xB2A5;&#xB825;&#xC744; &#xACB0;&#xD569; &#xC774;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xC2E0;&#xC18D;&#xD55C; &#xC720;&#xB3C4; &#xBC0F; &#xC2E4;&#xC2DC;&#xAC04;&#xC5D0;&#xC11C; &#xD615;&#xD0DC;&#xD559; &#xBC0F; &#xC140; &#xB3D9;&#xC791;&#xC5D0; &#xC601;&#xD5A5;&#xC744; &#xD3C9;&#xAC00; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xC778;&#xAC04;&#xC758; &#xC554; xenografts &#xB0B4;&#xC758; &#xB300;&#xC0C1; &#xB2E8;&#xBC31;&#xC9C8; &#xB808;&#xBCA8;&#xC758; &#xD6C4;&#xC18D; &#xC18C;&#xBAA8; &#xC788;&#xB294; intravital &#xC774;&#xBBF8;&#xC9D5; &#xBC29;&#xBC95;&#xC744; &#xAC1C;&#xBC1C;. &#xC870;&#xAC74;&#xC5D0; &#xB530;&#xB77C; &#xC548;&#xC815;&#xB41C; &#xD615;&#xAD11; &#xC804;&#xC790; cadherin &#xD0A4;&#xBA54;&#xB77C;&#xB294; &#xC804;&#xC774;&#xC131; &#xC720;&#xBC29;&#xC554; &#xC138;&#xD3EC;, &#xD45C;&#xD604;&#xACFC; &#xB1CC;&#xC885;&#xC591; &#xC870;&#xB958; &#xBC30;&#xC544; &#xBAA8;&#xB378;&#xC5D0; &#xC2E4;&#xC2DC;&#xAC04; confocal &#xD604;&#xBBF8;&#xACBD; &#xAC80;&#xC0AC; &#xBC95;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC804;&#xC790; cadherin &#xC720;&#xB3C4; &#xBC0F; &#xACE0;&#xAC08;&#xC758; &#xC601;&#xD5A5; &#xAC00;&#xC2DC;&#xD654; &#xD588;&#xB2E4;. &#xB2E8;&#xBC31;&#xC9C8; &#xC9C0; &#xBC29;&#xD654;, &#xC138;&#xD3EC; &#xD615;&#xD0DC;&#xD559; &#xBC0F; &#xC720; &#xBC29; &#xC885;&#xC591;&#xC5D0;&#xC11C; &#xAC04; &#xC5FD; &#xC0C1;&#xD53C;&#xC640; &#xC911;&#xAC04; &#xC5FD; &#xC0C1;&#xD53C; &#xC804;&#xD658;&#xC744; &#xACAA;&#xACE0; &#xC140;&#xC5D0; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158; &#xD3C9;&#xAC00; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;. &#xC774; &#xAE30;&#xC220;&#xC740; &#xC815;&#xBC00; &#xC81C;&#xC5B4; &#xAC00;&#xB2A5; &#xB2E8;&#xBC31;&#xC9C8; &#xD65C;&#xB3D9; vivo&#xC5D0;&#xC11C; &#xB3D9;&#xC801; &#xC0DD;&#xBB3C; &#xB9AC; &#xB9E4;&#xAC1C; &#xBCC0;&#xC218;&#xC758; &#xC2DC;&#xAC04;&#xC801; &#xBD84;&#xC11D;&#xC744; &#xD5C8;&#xC6A9; &#xD558;&#xB294; &#xB3D9;&#xC548;.

&#xC774;&#xBBF8;&#xC9D5; in vivo &#xC138;&#xD3EC; &#xC5ED;&#xD559;&#xC5D0; &#xD654;&#xD559;&#xC801;&#xC73C;&#xB85C; &#xC720;&#xB3C4;&#xD560; &#xC218; &#xC788;&#xB294; &#xB2E8;&#xBC31;&#xC9C8;&#xC758; &#xC601;&#xD5A5;.

El an&#xE1;lisis de eventos din&#xE1;micos en el microambiente del tumor durante la progresi&#xF3;n del c&#xE1;ncer est&#xE1; limitado por la complejidad de la corriente en modelos de im&#xE1;genes in vivo. A esto se suma la incapacidad de modular r&#xE1;pidamente y visualizar la actividad de la prote&#xED;na en tiempo real y de entender las consecuencias de estas perturbaciones en vivo. Hemos desarrollado un enfoque de imagen intravital que permite la r&#xE1;pida inducci&#xF3;n y el consiguiente agotamiento de los niveles de prote&#xED;na diana en xenoinjertos de c&#xE1;ncer de humanos, mientras que la evaluaci&#xF3;n del impacto en el comportamiento y la morfolog&#xED;a celular en tiempo real. Un condicional estabilizado fluorescentes E-cadherina quimera se expresa en las c&#xE9;lulas metast&#xE1;sicas del c&#xE1;ncer de mama, y &#x200B;&#x200B;el impacto de la inducci&#xF3;n de la E-cadherina y el agotamiento se visualizan en tiempo real mediante microscop&#xED;a confocal en un modelo de embri&#xF3;n aviar xenoinjerto. Se demuestra la evaluaci&#xF3;n de la localizaci&#xF3;n de la prote&#xED;na, la morfolog&#xED;a celular y la migraci&#xF3;n de las c&#xE9;lulas sometidas a la transici&#xF3;n epitelio-mes&#xE9;nquima y mesenquimal epitelial en los tumores de mama. Esta t&#xE9;cnica permite un control preciso sobre la actividad de la prote&#xED;na in vivo, al tiempo que permite el an&#xE1;lisis temporal de la din&#xE1;mica de los par&#xE1;metros biof&#xED;sicos.

Imagen del impacto de las prote&#xED;nas inducibles qu&#xED;micamente sobre la Din&#xE1;mica celular in vivo

Kisspeptins (KP), peptide products of the KISS1 metastasis-suppressor gene, are endogenous ligands for a G protein-coupled receptor (KISS1R). KISS1 acts as a metastasis suppressor in numerous human cancers. However, recent studies have demonstrated that an increase in KISS1 and KISS1R expression in patient breast tumors correlates with higher tumor grade and metastatic potential. We have shown that KP-10 stimulates invasion of estrogen receptor alpha (ERÎ±)-negative MDA-MB-231 breast cancer cells via transactivation of the epidermal growth factor receptor (EGFR). Here, we report that either KP-10 treatment of ERÎ±-negative non-malignant mammary epithelial MCF10A cells or expression of KISS1R in MCF10A cells induced a mesenchymal phenotype and stimulated invasiveness. Similarly, exogenous expression of KISS1R in ERÎ±-negative SKBR3 breast cancer cells was sufficient to trigger invasion and induced extravasation in vivo. In contrast, KP-10 failed to transactivate EGFR or stimulate invasiveness in the ERÎ±-positive MCF7 and T47D breast cancer cells. This suggested that ERÎ± negatively regulates KISS1R-dependent breast cancer cell migration, invasion and EGFR transactivation. In support of this, we found that these KP-10-induced effects were ablated upon exogenous expression of ERÎ± in the MDA-MB-231 cells, by downregulating KISS1R expression. Lastly, we have identified IQGAP1, an actin cytoskeletal binding protein as a novel binding partner of KISS1R, and have shown that KISS1R regulates EGFR transactivation in breast cancer cells in an IQGAP1-dependent manner. Overall, our data strongly suggest that the ERÎ± status of mammary cells dictates whether KISS1R may be a novel clinical target for treating breast cancer metastasis.

KISS1R Induces Invasiveness of Estrogen Receptor-Negative Human Mammary Epithelial and Breast Cancer Cells.

Kisspeptins (KP), produits de peptide du g&egrave;ne KISS1 m&eacute;tastase-suppresseur, sont des ligands endog&egrave;nes pour un r&eacute;cepteur coupl&eacute; aux prot&eacute;ines de G (KISS1R). KISS1 agit comme un suppresseur de m&eacute;tastase dans nombreux cancers humains. Toutefois, des &eacute;tudes r&eacute;centes ont d&eacute;montr&eacute; qu&#39;une augmentation de l&#39;expression KISS1 et KISS1R dans les tumeurs du sein patients est en corr&eacute;lation avec plus haut grade tumoral et le potentiel m&eacute;tastatique. Nous avons montr&eacute; que KP-10 stimule l&#39;invasion des &oelig;strog&egrave;nes alpha du r&eacute;cepteur (ER&alpha;)-n&eacute;gatif des cellules MDA-MB-231 mammaires canc&eacute;reuses par l&#39;interm&eacute;diaire de transactivation du r&eacute;cepteur du facteur de croissance &eacute;pidermique (EGFR). Nous rapportons ici, que soit KP-10 traitement des n&eacute;gatifs ER&alpha; b&eacute;nignes cellules MCF10A &eacute;pith&eacute;liales mammaires ou l&#39;expression de KISS1R dans les cellules de le MCF10A induite par un ph&eacute;notype m&eacute;senchymateux et stimul&eacute;e par des techniques invasives. De m&ecirc;me, expression exog&egrave;ne de KISS1R en cellules de cancer du sein SKBR3 ER&alpha; n&eacute;gatif &eacute;tait suffisante pour invasion de d&eacute;clencheur et extravasation induite in vivo. En revanche, KP-10 n&#39;a pas transactiver EGFR ou stimuler le pouvoir invasif des cellules canc&eacute;reuses du sein MCF7 ER&alpha; positifs et T47D. Cela sugg&egrave;re que ER&alpha; r&eacute;gule n&eacute;gativement KISS1R-d&eacute;pendante du sein cancer cell migration, l&#39;invasion et la transactivation EGFR. &Agrave; l&#39;appui de cela, nous avons constat&eacute; que ces effets induits par le KP-10 ont pratiqu&eacute; l&#39;ablation une expression exog&egrave;ne de ER&alpha; dans les cellules MDA-MB-231, par leur expression KISS1R. Enfin, nous avons identifi&eacute; IQGAP1, une prot&eacute;ine de liaison du cytosquelette d&#39;actine comme un partenaire de liaison roman de KISS1R et ont montr&eacute; que KISS1R r&eacute;gule la transactivation EGFR dans les cellules canc&eacute;reuses du sein de mani&egrave;re d&eacute;pendante IQGAP1. Dans l&#39;ensemble, nos r&eacute;sultats sugg&egrave;rent fortement que le statut ER&alpha; des cellules mammaires dicte si KISS1R peut &ecirc;tre une nouvelle cible clinique pour le traitement des m&eacute;tastases du cancer du sein.

KISS1R induit des techniques invasives d&#39;oestrog&egrave;ne r&eacute;cepteur humain mammaire &eacute;pith&eacute;liales et les cellules de Cancer des seins.

Kisspeptins (KP), prodotti del peptide del gene soppressore di metastasi KISS1, sono ligandi endogeni per un recettore accoppiati a proteine G (KISS1R). KISS1 agisce come un soppressore di metastasi in numerosi tumori umani. Tuttavia, recenti studi hanno dimostrato che un aumento nell&#39;espressione di KISS1 e di KISS1R in tumori al seno paziente correla con il grado pi&ugrave; elevato di tumore e potenziale metastatico. Abbiamo dimostrato che il KP-10 stimola invasione dell&#39;alfa del ricevitore dell&#39;estrogeno (ER&alpha;)-negativo cellule di cancro al seno MDA-MB-231 via transattivazione del recettore dell&#39;epidermal growth factor (EGFR). Qui, segnaliamo che KP-10 trattamento di ER&alpha;-negativo non maligne mammarie MCF10A cellule epiteliali o espressione di KISS1R nelle cellule MCF10A indotta da un fenotipo mesenchimale e stimolato invasivit&agrave;. Allo stesso modo, espressione esogena di KISS1R in cellule di cancro al seno negativo ER&alpha; SKBR3 era sufficiente per invasione di trigger e indotto stravaso in vivo. Al contrario, KP-10 non &egrave; riuscito a transactivate EGFR o stimolare invasivit&agrave; nelle cellule di cancro al seno MCF7 ER&alpha;-positivo e T47D. Questo ha suggerito che ER&alpha; regola negativamente KISS1R-dipendente al seno cancro cellula migrazione, invasione e transactivation EGFR. A sostegno di ci&ograve;, abbiamo trovato che questi effetti indotta da KP-10 erano ablazione su espressione esogena di ER&alpha; nelle cellule MDA-MB-231, da riducono espressione KISS1R. Infine, ci hanno identificato IQGAP1, una proteina actina del citoscheletro come partner associazione romanzo di KISS1R e hanno dimostrato che KISS1R regola la transattivazione di EGFR nelle cellule tumorali del seno in maniera IQGAP1-dipendente. Nel complesso, i nostri dati suggeriscono fortemente che lo status ER&alpha; delle cellule mammarie determina se KISS1R pu&ograve; essere un romanzo bersaglio clinico per il trattamento delle metastasi del cancro al seno.

KISS1R induce invasivit&agrave; dell&#39;estrogeno recettore-negativi umani mammario epiteliali e le cellule di cancro della mammella.

The combination of nanoparticle (NP) size, charge, and surface chemistry (e.g., extent of modification with polyethylene glycol (PEG)) is accepted as a key determinant of NP/cellular interactions. However, the influence of spatial arrangement and accessibility of the charged molecules on the NP surface vis-à-vis the average surface charge (zeta (ζ) potential) is incompletely understood. Here we demonstrate that two types of mesoporous silica nanoparticles (MSNP) that are matched in terms of primary and hydrodynamic particle size, shape, pore structure, colloidal stability, and ζ potential, but differ in surface chemistry, viz. the spatial arrangement and relative exposure of surface amines, have profoundly different interactions with cells and tissues when evaluated in vitro and in vivo. While both particles are ∼50 nm in diameter, PEGylated, and positively charged (ζ = +40 mV), PEG-PEI (MSNPs modified with exposed polyamines), but not PEG-NMe3(+) (MSNP modified with distributed, obstructed amines) rapidly bind serum proteins, diverse cells types in vitro, and endothelial and white blood cells in vivo (ex ovo chick embryo model). This finding helps elucidate the relative role of surface exposure of charged molecules vs ζ potential in otherwise physicochemically matched MSNP and highlights protein corona neutrality as an important design consideration when synthesizing cationic NPs for biological applications.

Re-examining the size/charge paradigm: differing in vivo characteristics of size- and charge-matched mesoporous silica nanoparticles.

Hypoxia promotes tumor cell motility via RhoA and ROCK1 signaling pathways.

Tumor heterogeneity confounds cancer diagnosis and the outcome of therapy, necessitating analysis of tumor cell subsets within the tumor mass. Elevated expression of hyaluronan (HA) and HA receptors, receptor for HA-mediated motility (RHAMM)/HA-mediated motility receptor and cluster designation 44 (CD44), in breast tumors correlates with poor outcome. We hypothesized that a probe for detecting HA-HA receptor interactions may reveal breast cancer (BCa) cell heterogeneity relevant to tumor progression. A fluorescent HA (F-HA) probe containing a mixture of polymer sizes typical of tumor microenvironments (10-480 kDa), multiplexed profiling, and flow cytometry were used to monitor HA binding to BCa cell lines of different molecular subtypes. Formulae were developed to quantify binding heterogeneity and to measure invasion in vivo. Two subsets exhibiting differential binding (HA(-/low) vs. HA(high)) were isolated and characterized for morphology, growth, and invasion in culture and as xenografts in vivo. F-HA-binding amounts and degree of heterogeneity varied with BCa subtype, were highest in the malignant basal-like cell lines, and decreased upon reversion to a nonmalignant phenotype. Binding amounts correlated with CD44 and RHAMM displayed but binding heterogeneity appeared to arise from a differential ability of HA receptor-positive subpopulations to interact with F-HA. HA(high) subpopulations exhibited significantly higher local invasion and lung micrometastases but, unexpectedly, lower proliferation than either unsorted parental cells or the HA(-/low) subpopulation. Querying F-HA binding to aggressive tumor cells reveals a previously undetected form of heterogeneity that predicts invasive/metastatic behavior and that may aid both early identification of cancer patients susceptible to metastasis, and detection/therapy of invasive BCa subpopulations.

Cellular heterogeneity profiling by hyaluronan probes reveals an invasive but slow-growing breast tumor subset.

Tumor cells have unstable genomes relative to non-tumor cells. Decreased DNA integrity resulting from tumor cell instability is important in generating favorable therapeutic indices, and intact DNA repair mediates resistance to therapy. Targeting DNA repair to promote the action of anti-cancer agents is therefore an attractive therapeutic strategy. BRCA2 is involved in homologous recombination repair. BRCA2 defects increase cancer risk but, paradoxically, cancer patients with BRCA2 mutations have better survival rates. We queried TCGA data and found that BRCA2 alterations led to increased survival in patients with ovarian and endometrial cancer. We developed a BRCA2-targeting second-generation antisense oligonucleotide (ASO), which sensitized human lung, ovarian, and breast cancer cells to cisplatin by as much as 60%. BRCA2 ASO treatment overcame acquired cisplatin resistance in head and neck cancer cells, but induced minimal cisplatin sensitivity in non-tumor cells. BRCA2 ASO plus cisplatin reduced respiration as an early event preceding cell death, concurrent with increased glucose uptake without a difference in glycolysis. BRCA2 ASO and cisplatin decreased metastatic frequency in vivo by 77%. These results implicate BRCA2 as a regulator of metastatic frequency and cellular metabolic response following cisplatin treatment. BRCA2 ASO, in combination with cisplatin, is a potential therapeutic anti-cancer agent.

BRCA2 inhibition enhances cisplatin-mediated alterations in tumor cell proliferation, metabolism, and metastasis.

Tumor cell extravasation is a key step during cancer metastasis, yet the precise mechanisms that regulate this dynamic process are unclear. We utilized a high-resolution time-lapse intravital imaging approach to visualize the dynamics of cancer cell extravasation in vivo. During intravascular migration, cancer cells form protrusive structures identified as invadopodia by their enrichment of MT1-MMP, cortactin, Tks4, and importantly Tks5, which localizes exclusively to invadopodia. Cancer cells extend invadopodia through the endothelium into the extravascular stroma prior to their extravasation at endothelial junctions. Genetic or pharmacological inhibition of invadopodia initiation (cortactin), maturation (Tks5), or function (Tks4) resulted in an abrogation of cancer cell extravasation and metastatic colony formation in an experimental mouse lung metastasis model. This provides direct evidence of a functional role for invadopodia during cancer cell extravasation and distant metastasis and reveals an opportunity for therapeutic intervention in this clinically important process.

Invadopodia are required for cancer cell extravasation and are a therapeutic target for metastasis.

Planar cell polarity (PCP) signaling has been shown in different studies to either promote or inhibit the malignancy of breast cancer. Using the 21T cell lines, which were derived from an individual patient and represent distinct stages of progression, we show that the prototypical PCP ligand, WNT5A, is expressed highest in 21MT-1 cells (invasive mammary carcinoma) and lowest in 21PT (atypical ductal hyperplasia) and 21NT (ductal carcinoma in situ) cells. Overexpression of WNT5A decreased spherical colony formation and increased invasion and in vivo extravasation only in 21NT cells; whereas overexpression increased migration of both 21PT and 21NT cells. WNT5A overexpression also increased RHOA expression of both cell lines and subsequent RHOA knockdown blocked WNT5A-induced migration, but only partially blocked WNT5A-induced invasion of 21NT cells. PCP can signal through VANGL1 to modulate AP-1 target genes (e.g. MMP3) and induce invasion. VANGL1 knockdown inhibited WNT5A-induced invasion of 21NT cells, but had no effect on WNT5A-induced migration of either 21PT or 21NT cells. WNT5A-induced MMP3 expression was seen only in 21NT cells, an effect that was VANGL1 dependent, but independent of AP-1. We thus provide evidence that PCP signaling can act in a context dependent manner to promote breast cancer progression.

Article	Year
Gender and post-ischemic recovery of hypertrophied rat hearts. BMC cardiovascular disorders\| PubMed ID: 16509993	2006
Exercise increases tissue-type plasminogen activator expression in rat cardiomyocytes. Thrombosis and haemostasis\| PubMed ID: 17139388	2006
Human 21T breast epithelial cell lines mimic breast cancer progression in vivo and in vitro and show stage-specific gene expression patterns. Laboratory investigation; a journal of technical methods and pathology\| PubMed ID: 20458274	2010
Imaging the impact of chemically inducible proteins on cellular dynamics in vivo. PloS one\| PubMed ID: 22276156	2012
KISS1R Induces Invasiveness of Estrogen Receptor-Negative Human Mammary Epithelial and Breast Cancer Cells. Endocrinology\| PubMed ID: 23525242	2013
Re-examining the size/charge paradigm: differing in vivo characteristics of size- and charge-matched mesoporous silica nanoparticles. Journal of the American Chemical Society\| PubMed ID: 24107191	2013
Hypoxia promotes tumor cell motility via RhoA and ROCK1 signaling pathways. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 24398519	2014
Cellular heterogeneity profiling by hyaluronan probes reveals an invasive but slow-growing breast tumor subset. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 24733940	2014
BRCA2 inhibition enhances cisplatin-mediated alterations in tumor cell proliferation, metabolism, and metastasis. Molecular oncology\| PubMed ID: 24974076	2014
Invadopodia are required for cancer cell extravasation and are a therapeutic target for metastasis. Cell reports\| PubMed ID: 25176655	2014
Stage of breast cancer progression influences cellular response to activation of the WNT/planar cell polarity pathway. Scientific reports\| PubMed ID: 25204426	2014
CORRIGENDUM: Stage of Breast Cancer Progression Influences Cellular Response to Activation of the WNT/Planar Cell Polarity Pathway. Scientific reports\| PubMed ID: 25613050	2015

Hon S. Leong

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