detection-human-leukocyte-antigen-biomarkers-breast-cancer-utilizing

Detection of Human Leukocyte Antigen Biomarkers in Breast Cancer Utilizing Label-free Biosensor Technology

According to the American Cancer Society, more than 200,000 women will be diagnosed with invasive breast cancer each year and approximately 40,000 will ...

Texas Tech University Health Sciences Center

Cellular immune mechanisms detect and destroy cancerous and infected cells via the human leukocyte antigen (HLA) class I molecules that present peptides of intracellular origin on the surface of all nucleated cells. The identification of novel, tumor-specific epitopes is a critical step in the development of immunotherapeutics for breast cancer. To directly identify peptide epitopes unique to cancerous cells, secreted human class I HLA molecules (sHLA) were constructed by deletion of the transmembrane and cytoplasmic domain of HLA A*0201. The resulting sHLA-A*0201 was transferred and expressed in breast cancer cell lines MCF-7, MDA-MB-231, and BT-20 as well as in the immortal, nontumorigenic cell line MCF10A. Stable transfectants were seeded into bioreactors for production of > 25 mg of sHLA-A*0201. Peptides eluted from affinity purified sHLA were analyzed by mass spectroscopy. Comparative analysis of HLA-A*0201 peptides revealed 5 previously uncharacterized epitopes uniquely presented on breast cancer cells. These peptides were derived from intracellular proteins with either well-defined or putative roles in breast cancer development and progression: Cyclin Dependent Kinase 2 (Cdk2), Ornithine Decarboxylase (ODC1), Kinetochore Associated 2 (KNTC2 or HEC1), Macrophage Migration Inhibitory Factor (MIF), and Exosome Component 6 (EXOSC6). Cellular recognition of the MIF, KNTC2, EXOSC6, and Cdk2 peptides by circulating CD8+ cells was demonstrated by tetramer staining and IFN-gamma ELISPOT. The identification and characterization of peptides unique to the class I of breast cancer cells provide putative targets for the development of immune diagnostic tools and therapeutics.

Identification of breast cancer peptide epitopes presented by HLA-A*0201.

HLA-B27- and -B57-positive HIV-infected humans have long been associated with control of HIV replication, implying that CD8(+) T cell responses contribute to control of viral replication. In a similar fashion, 50% of Mamu-B*08-positive Indian rhesus macaques control SIVmac239 replication and become elite controllers with chronic-phase viremia <1000 viral RNA copies/ml. Interestingly, Mamu-B*08-restricted SIV-derived epitopes appeared to match the peptide binding profile for HLA-B*2705 in humans. We therefore defined a detailed peptide-binding motif for Mamu-B*08 and investigated binding similarities between the macaque and human MHC class I molecules. Analysis of a panel of approximately 900 peptides revealed that despite substantial sequence differences between Mamu-B*08 and HLA-B*2705, the peptide-binding repertoires of these two MHC class I molecules share a remarkable degree of overlap. Detailed knowledge of the Mamu-B*08 peptide-binding motif enabled us to identify six additional novel Mamu-B*08-restricted SIV-specific CD8(+) T cell immune responses directed against epitopes in Gag, Vpr, and Env. All 13 Mamu-B*08-restricted epitopes contain an R at the position 2 primary anchor and 10 also possess either R or K at the N terminus. Such dibasic peptides are less prone to cellular degradation. This work highlights the relevance of the Mamu-B*08-positive SIV-infected Indian rhesus macaque as a model to examine elite control of immunodeficiency virus replication. The remarkable similarity of the peptide-binding motifs and repertoires for Mamu-B*08 and HLA-B*2705 suggests that the nature of the peptide bound by the MHC class I molecule may play an important role in control of immunodeficiency virus replication.

Two MHC class I molecules associated with elite control of immunodeficiency virus replication, Mamu-B*08 and HLA-B*2705, bind peptides with sequence similarity.

The identification and validation of new cancer-specific T cell epitopes continues to be a major area of research interest. Nevertheless, challenges remain to develop strategies that can easily discover and validate epitopes expressed in primary cancer cells. Regarded as targets for T cells, peptides presented in the context of the major histocompatibility complex (MHC) are recognized by monoclonal antibodies (mAbs). These mAbs are of special importance as they lend themselves to the detection of epitopes expressed in primary tumor cells. Here, we use an approach that has been successfully utilized in two different infectious disease applications (WNV and influenza). A direct peptide-epitope discovery strategy involving mass spectrometric analysis led to the identification of peptide YLLPAIVHI in the context of MHC A*02 allele (YLL/A2) from human breast carcinoma cell lines. We then generated and characterized an anti-YLL/A2 mAb designated as RL6A TCRm. Subsequently, the TCRm mAb was used to directly validate YLL/A2 epitope expression in human breast cancer tissue, but not in normal control breast tissue. Moreover, mice implanted with human breast cancer cells grew tumors, yet when treated with RL6A TCRm showed a marked reduction in tumor size. These data demonstrate for the first time a coordinated direct discovery and validation strategy that identified a peptide/MHC complex on primary tumor cells for antibody targeting and provide a novel approach to cancer immunotherapy.

Direct discovery and validation of a peptide/MHC epitope expressed in primary human breast cancer cells using a TCRm monoclonal antibody with profound antitumor properties.

MHC class I proteins are partly responsible for shaping the magnitude and focus of the adaptive cellular immune response. In humans, conventional wisdom suggests that the HLA-A, -B, and -C alleles are equally expressed on the majority of cell types. While we currently have a thorough understanding of how total MHC class I expression varies in different tissues, it has been difficult to examine expression of single MHC class I alleles due to the homogeneity of MHC class I sequences. It is unclear how cDNA species are expressed in distinct cell subsets in humans and particularly in macaques which transcribe upwards of 20 distinct MHC class I alleles at variable levels.

Differential MHC class I expression in distinct leukocyte subsets.

The contribution of CXCR7 to the tumor microenvironment has introduced a new level of complexity to CXCL12 signaling in breast cancer. In the previous issue of Breast Cancer Research, Hernandez and colleagues delineate the roles of CXCR4 and CXCR7 in tumor invasion and metastasis. The authors demonstrate that co-expression of CXCR7 and CXCR4 results in inhibition of CXCL12-mediated invasion, reduced intravasation of tumor cells into the vasculature, and fewer lung metastases compared with parental tumors. The results of this study suggest the combination of small molecule inhibitors of CXCR4 and CXCR7 could dramatically reduce invasion, intravasation, and metastasis and could be highly beneficial for the treatment of invasive breast cancer.

The dynamic yin-yang interaction of CXCR4 and CXCR7 in breast cancer metastasis.

The purpose of this preclinical study was to determine the effectiveness of RAF265, a multikinase inhibitor, for treatment of human metastatic melanoma and to characterize traits associated with drug response.

RAF265 inhibits the growth of advanced human melanoma tumors.

Although human angiosarcoma has been associated frequently with mutational inactivation of the tumor suppressor gene Ink4a/Arf, the underlying mechanisms have not been delineated. Here we report that malignant angiosarcoma is associated with high levels of RelA/NF-κB and IL-6 in contrast to normal vessels or benign hemagiomas. Studies of Ink4a/Arf deficient mice not only recapitulate genetic traits observed in human angiosarcoma, but also unveil a possible therapeutic link comprised of the NF-kB/IL-6/Stat3 signaling axis. In Ink4a/Arf(-/-) cells, NF-κB controlled Stat3 signaling by transcriptionally controlling the expression of IL-6, gp130, and Jak2. Further, IL-6 mediated Stat3 signaling through the sIL-6R. Inhibition of Ikkβ solely in myeloid cells was insufficient to block angiosarcoma development; in contrast, systemic inhibition of Ikkβ, IL-6, or Stat3 markedly inhibited angiosarcoma growth. Our findings offer clinical implications for targeting the NF-kB/IL-6/STAT3 pathway as a rational strategy to treat angiosarcoma.

INK4a/ARF [corrected] inactivation with activation of the NF-κB/IL-6 pathway is sufficient to drive the development and growth of angiosarcoma.

&#x764C;&#x57FA;&#x56E0;&#x8BF1;&#x5BFC;&#x8870;&#x8001;&#x53EF;&#x4EE5;&#x63D0;&#x4F9B;&#x9488;&#x5BF9;&#x80BF;&#x7624;&#x8FDB;&#x5C55;&#x7684;&#x4FDD;&#x62A4;&#x673A;&#x5236;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x7531;&#x8870;&#x8001;&#x7EC6;&#x80DE;&#x751F;&#x957F;&#x56E0;&#x5B50;&#x53CA;&#x7EC6;&#x80DE;&#x56E0;&#x5B50;&#x7684;&#x751F;&#x4EA7;&#x53EF;&#x80FD;&#x80BF;&#x7624;&#x53D1;&#x5C55;&#x4F5C;&#x51FA;&#x8D21;&#x732E;&#x3002;&#x56E0;&#x6B64;&#xFF0C;&#x4E0D;&#x6E05;&#x695A;&#x662F;&#x5426;&#x8870;&#x8001;&#x7684;&#x8BF1;&#x5BFC;&#x8868;&#x793A;&#x4E00;&#x79CD;&#x53EF;&#x884C;&#x7684;&#x6CBB;&#x7597;&#x65B9;&#x6CD5;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x4F7F;&#x7528;&#x53D6;&#x81EA; 19 &#x4F8B;&#x8F6C;&#x79FB;&#x6027;&#x9ED1;&#x8272;&#x7D20;&#x7624;&#x80BF;&#x7624;&#x539F;&#x4F4D;&#x690D;&#x5165;&#x5C0F;&#x9F20;&#x6A21;&#x578B;&#xFF0C;&#x6211;&#x4EEC;&#x6307;&#x51FA;&#x9488;&#x5BF9;&#x6781;&#x5149;&#x6FC0;&#x9176; MLN8054/MLN8237 &#x4E0E;&#x53D7;&#x635F;&#x7EC6;&#x80DE;&#x6709;&#x4E1D;&#x5206;&#x88C2;&#x3001; &#x8BF1;&#x5BFC;&#x8870;&#x8001;&#x663E;&#x8457;&#x963B;&#x6B62;&#x5728;&#x690D;&#x5165;&#x75C5;&#x4EBA;&#x7684;&#x80BF;&#x7624;&#x6269;&#x6563;&#x3002;&#x91CD;&#x8981;&#x7684;&#x662F;&#xFF0C;&#x5F53;&#x7624;&#x5C0F;&#x9F20;&#x7684;&#x5B50;&#x96C6;&#x88AB;&#x76D1;&#x6D4B;&#x80BF;&#x7624;&#x8FDB;&#x5C55;&#x540E; 12 &#x4E2A;&#x6708;&#x671F;&#x95F4;&#x6682;&#x505C; MLN8054 &#x6CBB;&#x7597;&#xFF0C;50%&#x7684;&#x80BF;&#x7624;&#x6CA1;&#x6709;&#x8FDB;&#x5C55;&#x3002;&#x673A;&#x68B0;&#x5206;&#x6790;&#x63ED;&#x793A;&#x7684;&#x6781;&#x5149;&#x6FC0;&#x9176;&#x6291;&#x5236;&#x8BF1;&#x5BFC;&#x591A;&#x500D;&#x4F53;&#x548C; ATM/Chk2 DNA &#x635F;&#x4F24;&#x54CD;&#x5E94;&#xFF0C;&#x4ECE;&#x800C;&#x4ECB;&#x5BFC;&#x7684;&#x8870;&#x8001;&#x548C; NF-&kappa; B &#x76F8;&#x5173;&#xFF0C;&#x8870;&#x8001;&#x76F8;&#x5173;&#x5206;&#x6CCC;&#x578B; &#xFF08;&#x67F3;&#x6C2E;&#x78FA;&#x5421;&#x5576;)&#x3002;IKK&beta; NF-&kappa; B &#x7684;&#x5C01;&#x9501;&#xFF0C;&#x5BFC;&#x81F4; MLN8237 &#x8BF1;&#x5BFC;&#x8870;&#x8001;&#x548C;&#x67F3;&#x6C2E;&#x78FA;&#x5421;&#x5576;&#x7684;&#x9006;&#x8F6C;&#x3002;&#x7ED3;&#x679C;&#x8868;&#x660E;&#x53BB;&#x9664;&#x8870;&#x8001;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x7684;&#x6D78;&#x6DA6;&#x9AD3;&#x7EC6;&#x80DE;&#x662F;&#x80BF;&#x7624;&#x518D;&#x751F;&#x957F;&#x7684;&#x6291;&#x5236;&#x4F5C;&#x7528;&#x7684;&#x5173;&#x952E;&#x3002;&#x603B;&#x4E4B;&#xFF0C;&#x8FD9;&#x4E9B;&#x6570;&#x636E;&#x8868;&#x660E;&#x8BF1;&#x5BFC;&#x7684;&#x8870;&#x8001;&#xFF0C;&#x52A0;&#x4E0A;&#x514D;&#x75AB;&#x76D1;&#x89C6;&#xFF0C;&#x53EF;&#x4EE5;&#x9650;&#x5236;&#x9ED1;&#x8272;&#x7D20;&#x7624;&#x751F;&#x957F;&#x3002;

&#x9488;&#x5BF9;&#x6781;&#x5149;&#x6FC0;&#x9176;&#x9650;&#x5236;&#x80BF;&#x7624;&#x751F;&#x957F;&#x901A;&#x8FC7; DNA &#x635F;&#x4F24;&#x4ECB;&#x5BFC;&#x8870;&#x8001;&#x548C; NF-&kappa; B &#x7684;&#x5C01;&#x9501;&#x635F;&#x5BB3;&#x6B64;&#x836F;&#x7269;&#x6240;&#x81F4;&#x7684;&#x8870;&#x8001;&#x3002;

Oncogene-induced senescence can provide a protective mechanism against tumour progression. However, production of cytokines and growth factors by senescent cells may contribute to tumour development. Thus, it is unclear whether induction of senescence represents a viable therapeutic approach. Here, using a mouse model with orthotopic implantation of metastatic melanoma tumours taken from 19 patients, we observed that targeting aurora kinases with MLN8054/MLN8237 impaired mitosis, induced senescence and markedly blocked proliferation in patient tumour implants. Importantly, when a subset of tumour-bearing mice were monitored for tumour progression after pausing MLN8054 treatment, 50% of the tumours did not progress over a 12-month period. Mechanistic analyses revealed that inhibition of aurora kinases induced polyploidy and the ATM/Chk2 DNA damage response, which mediated senescence and a NF-ÎºB-related, senescence-associated secretory phenotype (SASP). Blockade of IKKÎ²/NF-ÎºB led to reversal of MLN8237-induced senescence and SASP. Results demonstrate that removal of senescent tumour cells by infiltrating myeloid cells is crucial for inhibition of tumour re-growth. Altogether, these data demonstrate that induction of senescence, coupled with immune surveillance, can limit melanoma growth.

Targeting aurora kinases limits tumour growth through DNA damage-mediated senescence and blockade of NF-ÎºB impairs this drug-induced senescence.

S&eacute;nescence induite par l&#39;oncog&egrave;ne peut fournir un m&eacute;canisme de protection contre la progression tumorale. Toutefois, la production de cytokines et de facteurs de croissance de cellules s&eacute;nescentes pourrait contribuer au d&eacute;veloppement de la tumeur. Ainsi, on ignore si l&#39;induction de la s&eacute;nescence repr&eacute;sente une approche th&eacute;rapeutique viable. Ici, en utilisant un mod&egrave;le de souris avec implantation orthotopique de tumeurs de m&eacute;lanome m&eacute;tastatique pr&eacute;lev&eacute;es sur 19 patients, nous avons observ&eacute; que ciblant les kinases aurora avec MLN8054/MLN8237 avec facult&eacute;s affaiblies de la mitose, induit la s&eacute;nescence et bloqu&eacute; nettement la prolif&eacute;ration dans les tumeurs de patients implants. Quand ce qui est important, un sous-ensemble des souris porteuses de tumeurs ont &eacute;t&eacute; observ&eacute;es pendant la progression tumorale apr&egrave;s que suspension MLN8054 traitement, 50 % des tumeurs n&#39;a pas progress&eacute; sur une p&eacute;riode de 12 mois. M&eacute;canistes analyses ont r&eacute;v&eacute;l&eacute; que l&#39;inhibition des kinases aurora a induit polyplo&iuml;die et la r&eacute;ponse de dommages ATM/Chk2 ADN, qui v&eacute;hicul&eacute;e par la s&eacute;nescence et un NF-&kappa;B-associ&eacute;s, associ&eacute;s &agrave; la s&eacute;nescence ph&eacute;notype s&eacute;cr&eacute;toire (SASP). Blocus de IKK&beta;/NF-&kappa;B a conduit au renversement de la s&eacute;nescence induite par le MLN8237 et de la SASP. Les r&eacute;sultats d&eacute;montrent que pr&eacute;l&egrave;vement de cellules tumorales s&eacute;nescentes en infiltrant des cellules my&eacute;lo&iuml;des est crucial pour l&#39;inhibition de la re-croissance de la tumeur. Au total, ces r&eacute;sultats d&eacute;montrent que l&#39;induction de la s&eacute;nescence, coupl&eacute;e &agrave; la surveillance immunitaire, peut limiter la croissance de m&eacute;lanome.

Ciblage des kinases aurora limite la croissance de la tumeur par l&#39;interm&eacute;diaire de s&eacute;nescence m&eacute;di&eacute;e par les dommages de l&#39;ADN et blocus de NF-&kappa;B affaiblit cette s&eacute;nescence induite par le m&eacute;dicament.

Onkogen-induzierte Seneszenz kann ein Schutzmechanismus gegen Tumorprogression bereitstellen. Jedoch kann die Produktion von Zytokinen und Wachstumsfaktoren durch alternde Zellen zu Tumor-Entwicklung beitragen. Daher ist es unklar, ob Induktion von Seneszenz geeigneter therapeutischen Ansatz darstellt. Hier beobachtet mit einem Mausmodell mit Orthotopic Implantation von metastasierenden Melanoms Tumore 19 Patienten entnommen, wir, dass targeting Aurora Kinasen mit MLN8054/MLN8237 Mitose beeintr&auml;chtigt, Seneszenz induzierte und deutlich Proliferation im Patienten Tumor Implantate blockiert. Wichtig ist, wann eine Teilmenge von Tumor-tragenden M&auml;usen wurden &uuml;berwacht f&uuml;r Tumorprogression nach anhalten MLN8054 Behandlung, 50 % der Tumore &uuml;ber einen Zeitraum von 12 Monaten keine Fortschritte. Mechanistische Analyse ergab, dass Hemmung der Aurora-Kinasen Polyploidie induzierte und die ATM/Chk2 DNA-Schaden-Antwort, wodurch die Seneszenz und ein NF-&kappa;B-bezogenen vermittelt, Altern-verbundene sekretorische Ph&auml;notyp (SASP). Blockade von IKK&beta;/NF-&kappa;B f&uuml;hrte zur Umkehrung der MLN8237 induziert Seneszenz und SASP. Ergebnisse zeigen, dass die Entfernung der alternde Tumorzellen durch infiltrieren myeloide Zellen f&uuml;r Hemmung des Tumorwachstums R&uuml;ck-entscheidend ist. Insgesamt zeigen diese Daten, dass die Induktion von Seneszenz, gepaart mit immun-&Uuml;berwachung, Melanom Wachstum begrenzen kann.

Zielgruppenadressierung Aurora Kinasen begrenzt Tumorwachstum durch DNA-Sch&auml;den-vermittelte Seneszenz und Blockade des NF-&kappa;B beeintr&auml;chtigt diese Arzneimittel-induzierter Altern.

Senescenza oncogene-indotta pu&ograve; fornire un meccanismo protettivo contro la progressione del tumore. Tuttavia, produzione di citochine e fattori di crescita da cellule senescenti pu&ograve; contribuire allo sviluppo di tumore. Cos&igrave;, non &egrave; chiaro se l&#39;induzione della senescenza rappresenta un valido approccio terapeutico. Qui, utilizzando un modello del mouse con l&#39;impianto ortotopico di tumori melanoma metastatico prelevati da 19 pazienti, abbiamo osservato che targeting chinasi aurora con MLN8054/MLN8237 alterata mitosi, indotta senescenza e marcatamente bloccata proliferazione nel paziente tumorale impianti. Importante, quando un sottoinsieme del tumore-cuscinetto topi sono stati monitorati per progressione tumorale dopo la sospensione dell&#39;esecuzione MLN8054 trattamento, il 50% dei tumori non ha fatto progredire per un periodo di 12 mesi. Meccanicistiche analisi hanno rivelato che l&#39;inibizione della chinasi aurora indotta poliploidia e la risposta di danno ATM/Chk2 DNA, che ha mediato un NF-&kappa;B correlati e senescenza, senescenza-associato fenotipo secretorio (SASP). Blocco della IKK&beta;/NF-&kappa;B portato ad inversione di MLN8237-indotta senescenza e SASP. Risultati dimostrano che la rimozione delle cellule tumorali senescenti da infiltrazione di cellule mieloidi &egrave; cruciale per l&#39;inibizione della ricrescita tumorale. Complessivamente, questi dati dimostrano che induzione della senescenza, accoppiato con sorveglianza immunitaria, pu&ograve; limitare la crescita del melanoma.

Targeting chinasi aurora limita la crescita del tumore attraverso la senescenza del DNA mediata da danni e blocco di NF-&kappa;B altera questo farmaco-indotta senescenza.

&#x764C;&#x907A;&#x4F1D;&#x5B50;&#x304C;&#x8A98;&#x5C0E;&#x3059;&#x308B;&#x8001;&#x5316;&#x306F;&#x3001;&#x816B;&#x760D;&#x306E;&#x9032;&#x884C;&#x306B;&#x5BFE;&#x3059;&#x308B;&#x4FDD;&#x8B77;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3092;&#x63D0;&#x4F9B;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x305F;&#x3060;&#x3057;&#x3001;&#x8001;&#x5316;&#x7D30;&#x80DE;&#x5897;&#x6B96;&#x56E0;&#x5B50;&#x30FB; &#x30B5;&#x30A4;&#x30C8;&#x30AB;&#x30A4;&#x30F3;&#x816B;&#x760D;&#x306E;&#x958B;&#x767A;&#x306B;&#x8CA2;&#x732E;&#x3059;&#x308B;&#x304B;&#x3082;&#x3057;&#x308C;&#x306A;&#x3044;&#x3002;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;&#x305D;&#x308C;&#x8001;&#x5316;&#x306E;&#x8A98;&#x5C0E;&#x304C;&#x5B9F;&#x884C;&#x53EF;&#x80FD;&#x306A;&#x6CBB;&#x7642;&#x30A2;&#x30D7;&#x30ED;&#x30FC;&#x30C1;&#x3092;&#x8868;&#x3059;&#x304B;&#x3069;&#x3046;&#x304B;&#x660E;&#x78BA;&#x3067;&#x306F;&#x306A;&#x3044;&#x3067;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x30DE;&#x30A6;&#x30B9;&#x3092;&#x7528;&#x3044;&#x3066;&#x540C;&#x6240;&#x6027;&#x540C;&#x7A2E;&#x79FB;&#x690D; 19 &#x60A3;&#x8005;&#x304B;&#x3089;&#x63A1;&#x53D6;&#x3057;&#x305F;&#x8EE2;&#x79FB;&#x6027;&#x9ED2;&#x8272;&#x816B;&#x306E;&#x816B;&#x760D;&#x306E;&#x30AA;&#x30FC;&#x30ED;&#x30E9; &#x30AD;&#x30CA;&#x30FC;&#x30BC; MLN8054/MLN8237 &#x3068;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x6709;&#x7CF8;&#x5206;&#x88C2;&#x969C;&#x5BB3;&#x3001;&#x8001;&#x5316;&#x3092;&#x8A98;&#x5C0E;&#x3057;&#x3001;&#x8457;&#x3057;&#x304F;&#x5897;&#x6B96;&#x3067;&#x306F;&#x60A3;&#x8005;&#x306E;&#x816B;&#x760D;&#x30A4;&#x30F3;&#x30D7;&#x30E9;&#x30F3;&#x30C8; &#x30D6;&#x30ED;&#x30C3;&#x30AF;&#x89B3;&#x5BDF;&#x3002;&#x91CD;&#x8981;&#x306A;&#x306F;&#x3001;&#x3068;&#x304D;&#x30DE;&#x30A6;&#x30B9;&#x816B;&#x760D;&#x306E;&#x30B5;&#x30D6;&#x30BB;&#x30C3;&#x30C8;&#x304C;&#x30E2;&#x30CB;&#x30BF;&#x30FC;&#x816B;&#x760D;&#x9032;&#x884C;&#x3092;&#x4E00;&#x6642;&#x505C;&#x6B62; MLN8054 &#x306E;&#x6CBB;&#x7642;&#x3001;&#x816B;&#x760D;&#x306E; 50% &#x306F; 12 &#x30AB;&#x6708;&#x306E;&#x9593;&#x9032;&#x884C;&#x3057;&#x306A;&#x304B;&#x3063;&#x305F;&#x5F8C;&#x3002;&#x6A5F;&#x69CB;&#x89E3;&#x6790;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x305F;&#x30AA;&#x30FC;&#x30ED;&#x30E9; &#x30AD;&#x30CA;&#x30FC;&#x30BC;&#x306E;&#x6291;&#x5236;&#x306B;&#x3088;&#x308B;&#x500D;&#x6570;&#x6027;&#x3068;&#x5A92;&#x4ECB;&#x306E;&#x8001;&#x5316;&#x3068; NF &kappa;B &#x95A2;&#x9023;&#x3001;ATM/Chk2 DNA &#x640D;&#x50B7;&#x5FDC;&#x7B54;&#x95A2;&#x9023;&#x5206;&#x6CCC;&#x8868;&#x73FE;&#x578B; (SASP)&#x3002;IKK&beta;/NF-&kappa; MLN8237 &#x304C;&#x8A98;&#x5C0E;&#x3059;&#x308B;&#x8001;&#x5316;&#x3068; SASP &#x306E;&#x53CD;&#x8EE2;&#x306B;&#x3064;&#x306A;&#x304C;&#x3063;&#x305F;&#x306E;&#x5C01;&#x9396;&#x3002;&#x7D50;&#x679C;&#x306F;&#x9AA8;&#x9AC4;&#x7CFB;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x306B;&#x3088;&#x308B;&#x8001;&#x5316;&#x306E;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x306E;&#x9664;&#x53BB;&#x304C;&#x816B;&#x760D;&#x306E;&#x518D;&#x5897;&#x6B96;&#x306E;&#x6291;&#x5236;&#x306E;&#x305F;&#x3081;&#x306B;&#x91CD;&#x8981;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x5B8C;&#x5168;&#x306B;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x30C7;&#x30FC;&#x30BF;&#x306F;&#x3001;&#x8A98;&#x5C0E;&#x7D50;&#x5408;&#x514D;&#x75AB;&#x76E3;&#x8996;&#x3057;&#x3001;&#x8001;&#x5316;&#x306E;&#x60AA;&#x6027;&#x9ED2;&#x8272;&#x816B;&#x306E;&#x6210;&#x9577;&#x3092;&#x5236;&#x9650;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

&#x30AA;&#x30FC;&#x30ED;&#x30E9; &#x30FB; &#x30AD;&#x30CA;&#x30FC;&#x30BC; &#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8; DNA &#x640D;&#x50B7;&#x3092;&#x4ECB;&#x3057;&#x305F;&#x8001;&#x5316;&#x306B;&#x3088;&#x308B;&#x816B;&#x760D;&#x6210;&#x9577;&#x3092;&#x5236;&#x9650;&#x3057;&#x3001;&#x3053;&#x306E;&#x85AC;&#x7269;&#x8A98;&#x767A;&#x6027;&#x8001;&#x5316; NF-&kappa; &#x306E;&#x5C01;&#x9396;&#x3092;&#x640D;&#x306A;&#x3044;&#x307E;&#x3059;&#x3002;

&#xB178; &#xC1E0;&#xB97C; oncogene &#xC720;&#xB3C4; &#xB41C; &#xC885;&#xC591; &#xC9C4;&#xD589;&#xC5D0; &#xB300; &#xD55C; &#xBCF4;&#xD638; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC744; &#xC81C;&#xACF5;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, cytokines &#xBC0F; &#xB178;&#xD654; &#xC138;&#xD3EC;&#xC5D0; &#xC758;&#xD574; &#xC131;&#xC7A5; &#xC694;&#xC778;&#xC758; &#xC0DD;&#xC0B0; &#xC885;&#xC591; &#xAC1C;&#xBC1C;&#xC5D0; &#xAE30;&#xC5EC;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xB530;&#xB77C;&#xC11C;, &#xADF8;&#xAC83;&#xC740; &#xB178;&#xD654;&#xC758; &#xC720;&#xB3C4; &#xAC00;&#xB2A5;&#xD55C; &#xCE58;&#xB8CC; &#xBC29;&#xBC95;&#xC744; &#xB098;&#xD0C0;&#xB0B4;&#xB294;&#xC9C0; &#xC5EC;&#xBD80; &#xBD84;&#xBA85; &#xB418;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;, MLN8054/mln8237&#xC640; &#xC624;&#xB85C;&#xB77C; kinases &#xB300;&#xC0C1;&#xC73C;&#xB85C; &#xC720;&#xC0AC; &#xBD84;&#xC5F4; &#xC7A5;&#xC560;, &#xB178;&#xD654;&#xB97C; &#xC720;&#xBC1C;&#xC744; &#xD604;&#xC800; &#xD558; &#xAC8C; &#xC784; &#xD50C; &#xB780; &#xD2B8; &#xD658;&#xC790; &#xC885;&#xC591;&#xC758;&#xC5D0;&#xC11C; &#xD655;&#xC0B0;&#xC744; &#xCC28;&#xB2E8; &#xAD00;&#xCC30; 19 &#xD658;&#xC790;&#xC5D0;&#xC11C; &#xAC00;&#xC838;&#xC628; &#xC804;&#xC774;&#xC131; &#xD751;&#xC0C9; &#xC885; &#xC885;&#xC591;&#xC758; orthotopic &#xC8FC;&#xC785; &#xB41C; &#xB9C8;&#xC6B0;&#xC2A4; &#xBAA8;&#xB378;&#xC744; &#xC0AC;&#xC6A9; &#xD569;&#xB2C8;&#xB2E4;. &#xC911;&#xC694; &#xD55C; &#xAC83;&#xC740; &#xB54C; &#xC885;&#xC591; &#xBC29;&#xC704; &#xC950;&#xC758; &#xD558;&#xC704; &#xC9D1;&#xD569; &#xD588;&#xB2E4; &#xBAA8;&#xB2C8;&#xD130;&#xB9C1; &#xC885;&#xC591; &#xC9C4;&#xD589;&#xC5D0; &#xB300; &#xD55C; &#xD6C4; &#xC77C;&#xC2DC; &#xC911;&#xC9C0; MLN8054 &#xCE58;&#xB8CC;&#xB294; &#xC885;&#xC591;&#xC758; 50%&#xB294; 12 &#xAC1C;&#xC6D4;&#xC5D0; &#xAC78;&#xCCD0; &#xC9C4;&#xD589; &#xB418;&#xC9C0; &#xC54A;&#xC558;&#xB2E4;. &#xAE30;&#xACC4; &#xBD84;&#xC11D; &#xBC1D;&#xD600; &#xC624;&#xB85C;&#xB77C; kinases &#xC5B5;&#xC81C; &#xC720;&#xB3C4; polyploidy &#xB178;&#xD654;&#xC640;&#xB294; NF-&kappa;B &#xAD00;&#xB828; &#xC911;&#xC7AC;, ATM/Chk2 DNA &#xC190;&#xC0C1; &#xC751;&#xB2F5; &#xB178;&#xD654; &#xAD00;&#xB828; &#xB41C; &#xBD84; &#xD615; (SASP). IKK&beta;/NF-&kappa;B MLN8237-&#xC720;&#xB3C4; &#xB178;&#xD654; &#xBC0F; SASP&#xC758; &#xBC18;&#xC804;&#xC5D0;&#xC758; &#xBD09;&#xC1C4;. &#xACB0;&#xACFC; &#xACE8;&#xC218;&#xC131; &#xC138;&#xD3EC;&#xC5D0; &#xCE68;&#xD22C; &#xD558; &#xC5EC; &#xB178;&#xD654; &#xC885;&#xC591; &#xC138;&#xD3EC;&#xC758; &#xC81C;&#xAC70;&#xB294; &#xC885;&#xC591; &#xC7AC; &#xC131;&#xC7A5;&#xC758; &#xC800;&#xD574;&#xC5D0; &#xB300; &#xD55C; &#xC911;&#xC694; &#xD55C; &#xC785;&#xC99D;. &#xBAA8;&#xB450; &#xC774;&#xB7EC;&#xD55C; &#xB370;&#xC774;&#xD130; &#xBCF4;&#xC5EC; &#xC720;&#xB3C4; &#xB178;&#xD654;, &#xBA74;&#xC5ED; &#xAC10;&#xC2DC;&#xC640; &#xACB0;&#xD569;&#xC758; &#xD751;&#xC0C9; &#xC885; &#xC131;&#xC7A5;&#xC744; &#xC81C;&#xD55C;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;.

DNA &#xC190;&#xC0C1;&#xC744; &#xC911;&#xC7AC; &#xB178;&#xD654;&#xB97C; &#xD1B5;&#xD574; &#xC885;&#xC591; &#xC131;&#xC7A5;&#xC744; &#xC81C;&#xD55C; &#xB300;&#xC0C1;&#xC73C;&#xB85C; &#xC624;&#xB85C;&#xB77C; kinases&#xC640; NF-&Kappa;b&#xC758; &#xBD09;&#xC1C4; &#xC57D;&#xBB3C; &#xC720;&#xBC1C; &#xB178;&#xD654;&#xAC00; &#xC190;&#xC0C1;.

Senescencia inducida por el oncog&eacute;n puede proporcionar un mecanismo de protecci&oacute;n contra la progresi&oacute;n del tumor. Sin embargo, la producci&oacute;n de citoquinas y factores de crecimiento de las c&eacute;lulas senescentes puede contribuir al desarrollo tumoral. Por lo tanto, no est&aacute; claro si la inducci&oacute;n de senescencia representa un enfoque terap&eacute;utico viable. Aqu&iacute;, mediante un modelo de rat&oacute;n con implantaci&oacute;n ortot&oacute;pica de tumores de melanoma metast&aacute;sico de 19 pacientes, observamos que dirigidas a cinasas aurora con MLN8054/MLN8237 deterioraron mitosis, inducida por senescencia y marcado bloquearon la proliferaci&oacute;n en implantes del tumor del paciente. Lo importante es que, cuando un subconjunto de tumor-cojinete ratones fueron supervisados para la progresi&oacute;n del tumor despu&eacute;s de pausa MLN8054 tratamiento, 50% de los tumores no progresar durante un per&iacute;odo de 12 meses. An&aacute;lisis mecanicistas revelaron que la inhibici&oacute;n de las quinasas aurora inducida por poliploid&iacute;a y senescencia asociada a la respuesta de da&ntilde;o del ADN ATM/Chk2, que mediada por senescencia y un NF-&kappa;B-relacionados, fenotipo secretor (SASP). Bloqueo de IKK&beta;/NF-&kappa;B condujo a revocaci&oacute;n de senescencia inducida por MLN8237 y spas. Resultados demuestran que la eliminaci&oacute;n de las c&eacute;lulas senescentes tumor infiltrando en c&eacute;lulas mieloides es crucial para la inhibici&oacute;n del crecimiento tumoral. En conjunto, estos datos demuestran que la inducci&oacute;n de senescencia, juntada con la vigilancia inmune, puede limitar el crecimiento del melanoma.

Dirigidos a las quinasas aurora limita el crecimiento de tumor a trav&eacute;s de senescencia mediada por da&ntilde;o de ADN y bloqueo de NF-&kappa;B deteriora este senescencia inducida por medicamentos.

Bone marrow-derived human mesenchymal stem cells (hMSCs) either promote or inhibit cancer progression, depending on factors that heretofore have been undefined. Here we have utilized extreme hypoxia (0.5% O2) and concurrent treatment with metal carcinogen (nickel) to evaluate the passage-dependent response of hMSCs toward cancerous transformation. Effects of hypoxia and nickel treatment on hMSC proliferation, apoptosis, gene and protein expression, replicative senescence, reactive oxygen species (ROS), redox mechanisms, and in vivo tumor growth were analyzed. The behavior of late passage hMSCs in a carcinogenic hypoxia environment follows a profile similar to that of transformed cancer cells (i.e., increased expression of oncogenic proteins, decreased expression of tumor suppressor protein, increased proliferation, decreased apoptosis, and aberrant redox mechanisms), but this effect was not observed in earlier passage control cells. These events resulted in accumulated intracellular ROS in vitro and excessive proliferation in vivo. We suggest a mechanism by which carcinogenic hypoxia modulates the activity of three critical transcription factors (c-MYC, p53, and HIF1), resulting in accumulated ROS and causing hMSCs to undergo cancer-like behavioral changes. This is the first study to utilize carcinogenic hypoxia as an environmentally relevant experimental model for studying the age-dependent cancerous transformation of hMSCs.

Passage-dependent cancerous transformation of human mesenchymal stem cells under carcinogenic hypoxia.

Myeloid cells are capable of promoting or eradicating tumor cells and the nodal functions that contribute to their different roles are still obscure. Here, we show that mice with myeloid-specific genetic loss of the NF-κB pathway regulatory kinase IKKβ exhibit more rapid growth of cutaneous and lung melanoma tumors. In a BRAF(V600E/PTEN(-/-)) allograft model, IKKβ loss in macrophages reduced recruitment of myeloid cells into the tumor, lowered expression of MHC class II molecules, and enhanced production of the chemokine CCL11, thereby negatively regulating dendritic-cell maturation. Elevated serum and tissue levels of CCL11 mediated suppression of dendritic-cell differentiation/maturation within the tumor microenvironment, skewing it toward a Th2 immune response and impairing CD8(+) T cell-mediated tumor cell lysis. Depleting macrophages or CD8(+) T cells in mice with wild-type IKKβ myeloid cells enhanced tumor growth, where the myeloid cell response was used to mediate antitumor immunity against melanoma tumors (with less dependency on a CD8(+) T-cell response). In contrast, myeloid cells deficient in IKKβ were compromised in tumor cell lysis, based on their reduced ability to phagocytize and digest tumor cells. Thus, mice with continuous IKKβ signaling in myeloid-lineage cells (IKKβ(CA)) exhibited enhanced antitumor immunity and reduced melanoma outgrowth. Collectively, our results illuminate new mechanisms through which NF-κB signaling in myeloid cells promotes innate tumor surveillance.

Myeloid IKKβ promotes antitumor immunity by modulating CCL11 and the innate immune response.

Therapeutics that induce cancer cell senescence can block cell proliferation and promote immune rejection. However, the risk of tumor relapse due to senescence escape may remain high due to the long lifespan of senescent cells that are not cleared. Here, we show how combining a senescence-inducing inhibitor of the mitotic kinase Aurora A (AURKA) with an MDM2 antagonist activates p53 in senescent tumors harboring wild-type 53. In the model studied, this effect is accompanied by proliferation arrest, mitochondrial depolarization, apoptosis, and immune clearance of cancer cells by antitumor leukocytes in a manner reliant upon Ccl5, Ccl1, and Cxcl9. The AURKA/MDM2 combination therapy shows adequate bioavailability and low toxicity to the host. Moreover, the prominent response of patient-derived melanoma tumors to coadministered MDM2 and AURKA inhibitors offers a sound rationale for clinical evaluation. Taken together, our work provides a preclinical proof of concept for a combination treatment that leverages both senescence and immune surveillance to therapeutic ends.

Article	Year
Identification of breast cancer peptide epitopes presented by HLA-A*0201. Journal of proteome research\| PubMed ID: 18345606	2008
Two MHC class I molecules associated with elite control of immunodeficiency virus replication, Mamu-B08 and HLA-B2705, bind peptides with sequence similarity. Journal of immunology (Baltimore, Md. : 1950)\| PubMed ID: 19494300	2009
Direct discovery and validation of a peptide/MHC epitope expressed in primary human breast cancer cells using a TCRm monoclonal antibody with profound antitumor properties. Cancer immunology, immunotherapy : CII\| PubMed ID: 19779714	2010
Differential MHC class I expression in distinct leukocyte subsets. BMC immunology\| PubMed ID: 21762519	2011
The dynamic yin-yang interaction of CXCR4 and CXCR7 in breast cancer metastasis. Breast cancer research : BCR\| PubMed ID: 22293321	2012
RAF265 inhibits the growth of advanced human melanoma tumors. Clinical cancer research : an official journal of the American Association for Cancer Research\| PubMed ID: 22351689	2012
INK4a/ARF [corrected] inactivation with activation of the NF-κB/IL-6 pathway is sufficient to drive the development and growth of angiosarcoma. Cancer research\| PubMed ID: 22836752	2012
Targeting aurora kinases limits tumour growth through DNA damage-mediated senescence and blockade of NF-ÎºB impairs this drug-induced senescence. EMBO molecular medicine\| PubMed ID: 23180582	2013
Passage-dependent cancerous transformation of human mesenchymal stem cells under carcinogenic hypoxia. FASEB journal : official publication of the Federation of American Societies for Experimental Biology\| PubMed ID: 23568779	2013
Myeloid IKKβ promotes antitumor immunity by modulating CCL11 and the innate immune response. Cancer research\| PubMed ID: 25336190	2014
Mdm2 and aurora kinase a inhibitors synergize to block melanoma growth by driving apoptosis and immune clearance of tumor cells. Cancer research\| PubMed ID: 25398437	2015

Oriana E. Hawkins

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