Identification of resistance mechanism to targeting of the ADAM 17/ EGFR axis in triple negative breast cancer in vivo

Triple-negative breast cancers (TNBCs) (ER/PR/HER2 negative) represent 15% of invasive\nbreast cancers and occur at a higher rate in young and African-American women. Exploration of\nnovel therapeutic approaches is critical, since only 30% of woman with metastatic breast cancer\nwill survive and vir...

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Autor principal: Sharma, Neha
Otros Autores: Zirlik, Andreas
Formato: Tesis de maestría acceptedVersion
Lenguaje:Inglés
Publicado: Facultad de Farmacia y Bioquímica 2015
Materias:
In-Vitro
Cáncer
Cáncer de mama triple negativo
ADAM 17/ EGFR
Triple Negative Breast Cancer
Ciencia de la vida
Acceso en línea:http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_835
http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_835.dir/835.PDF
Aporte de:
Repositorio Digital de la Universidad de Buenos Aires (UBA) de Universidad de Buenos Aires
id I28-R145-HWA_835
record_format dspace
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-145
collection Repositorio Digital de la Universidad de Buenos Aires (UBA)
language Inglés
orig_language_str_mv eng
topic In-Vitro
Cáncer
Cáncer de mama triple negativo
ADAM 17/ EGFR
Triple Negative Breast Cancer
Ciencia de la vida
spellingShingle In-Vitro
Cáncer
Cáncer de mama triple negativo
ADAM 17/ EGFR
Triple Negative Breast Cancer
Ciencia de la vida
Sharma, Neha
Identification of resistance mechanism to targeting of the ADAM 17/ EGFR axis in triple negative breast cancer in vivo
topic_facet In-Vitro
Cáncer
Cáncer de mama triple negativo
ADAM 17/ EGFR
Triple Negative Breast Cancer
Ciencia de la vida
description Triple-negative breast cancers (TNBCs) (ER/PR/HER2 negative) represent 15% of invasive\nbreast cancers and occur at a higher rate in young and African-American women. Exploration of\nnovel therapeutic approaches is critical, since only 30% of woman with metastatic breast cancer\nwill survive and virtually none with metastatic TNBC. The status quo as it pertains to the\ntreatment of TNBCs can be summarized as: no effective therapies available. In part, the lack of\ntherapeutic success is due to high genetic heterogeneity of TNBCs, challenging single drug\napproaches.\nMany targeted strategies to treat TNBC are being explored, including the inhibition of kinase\npathways (e.g. PI3K/Akt, MEK, VEGFR and PDGFR), the inhibition of DNA repair, of survival\npathways and androgen receptor blockade. In most cases, such single-drug targeted therapy is\ncombined with systemic genotoxic chemotherapy. For example, although about 60% of basallike\nTNBCs over express EGFR, EGFR targeted therapy, including kinase inhibition, has been\ndisappointing due to the development of resistance.\nVarious resistance mechanisms allow cancer cells to evade single-drug targeted therapies:\nmutations in the targeted molecules, extensive crosstalk/pathway redundancy and the upregulation\nof alternate growth or survival pathways. Design of combinatorial approaches of\ntherapeutics for TNBC that overcome resistance is therefore critical. The contribution of the\nproposed research is expected to be the identification of signaling network perturbations that\noccur in response to single targeted therapies, in particular in ADAM17/EGFR axis inhibition,\nand confer resistance. Based on published results, inhibition of the ADAM17/EGFR ligand axis\nin TNBC should provide therapeutic benefit with reduced tumor growth and decreased\nmetastasis, if not possibly cure.\nIn our in vitro studies on PKC? and PPP1R14D gene knockout in MDA-MB-231 cells indeed led\nto decreases in cellular growth and migration. However, to our surprise, when the same cells\nwere injected into mice through orthotropic fat pad transplantation, they produced aggressive,\nmetastatic tumors that showed activation of alternate growth signaling pathways, namely of the\nmitogen-activated protein kinase ERK and of the PI3kinase target Akt, also a mitogen activated\nkinase. This suggested that TNBC cells were developing resistance to EGF ligand regulator\nknockdown by rewiring their growth factor signaling pathways. To determine where these\nadditional growth signals come from, we first considered the tumor cells themselves.\nIn this context we discovered that when kept in culture, MDA-MB-231 cells expressing sh-\nRNAs targeting either PKCa or PPP1R14D maintained knockdown of the target for up to 35days\ntested. At the same time EGFR and ERK showed low activity as expected due to a decrease in\nEGF ligand cleavage; Akt activity was undetectable. Since we observed strong reactivation of\nERK and new activation of Akt in tumors in vivo, we considered possible up-regulation of other\ngrowth factor receptors on the cell surface that would be engaged by factors released from the\ntumor stroma once cells are inserted in vivo. Indeed, we found that FGFR2 and Erbb4 were\nupregulated. It is therefore likely that reactivation of ERK and new activation of Akt was due to\nFGFR2 and Erbb4. This would suggest that combination therapy of EGF ligand release regulator\ninhibition and FGFR inhibition would decrease growth of these tumors in vivo.
author2 Zirlik, Andreas
author_facet Zirlik, Andreas
Sharma, Neha
format Tesis de maestría
Tesis de maestría
acceptedVersion
author Sharma, Neha
author_sort Sharma, Neha
title Identification of resistance mechanism to targeting of the ADAM 17/ EGFR axis in triple negative breast cancer in vivo
title_short Identification of resistance mechanism to targeting of the ADAM 17/ EGFR axis in triple negative breast cancer in vivo
title_full Identification of resistance mechanism to targeting of the ADAM 17/ EGFR axis in triple negative breast cancer in vivo
title_fullStr Identification of resistance mechanism to targeting of the ADAM 17/ EGFR axis in triple negative breast cancer in vivo
title_full_unstemmed Identification of resistance mechanism to targeting of the ADAM 17/ EGFR axis in triple negative breast cancer in vivo
title_sort identification of resistance mechanism to targeting of the adam 17/ egfr axis in triple negative breast cancer in vivo
publisher Facultad de Farmacia y Bioquímica
publishDate 2015
url http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_835
http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_835.dir/835.PDF
work_keys_str_mv AT sharmaneha identificationofresistancemechanismtotargetingoftheadam17egfraxisintriplenegativebreastcancerinvivo
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spelling I28-R145-HWA_8352019-09-27 Triple-negative breast cancers (TNBCs) (ER/PR/HER2 negative) represent 15% of invasive\nbreast cancers and occur at a higher rate in young and African-American women. Exploration of\nnovel therapeutic approaches is critical, since only 30% of woman with metastatic breast cancer\nwill survive and virtually none with metastatic TNBC. The status quo as it pertains to the\ntreatment of TNBCs can be summarized as: no effective therapies available. In part, the lack of\ntherapeutic success is due to high genetic heterogeneity of TNBCs, challenging single drug\napproaches.\nMany targeted strategies to treat TNBC are being explored, including the inhibition of kinase\npathways (e.g. PI3K/Akt, MEK, VEGFR and PDGFR), the inhibition of DNA repair, of survival\npathways and androgen receptor blockade. In most cases, such single-drug targeted therapy is\ncombined with systemic genotoxic chemotherapy. For example, although about 60% of basallike\nTNBCs over express EGFR, EGFR targeted therapy, including kinase inhibition, has been\ndisappointing due to the development of resistance.\nVarious resistance mechanisms allow cancer cells to evade single-drug targeted therapies:\nmutations in the targeted molecules, extensive crosstalk/pathway redundancy and the upregulation\nof alternate growth or survival pathways. Design of combinatorial approaches of\ntherapeutics for TNBC that overcome resistance is therefore critical. The contribution of the\nproposed research is expected to be the identification of signaling network perturbations that\noccur in response to single targeted therapies, in particular in ADAM17/EGFR axis inhibition,\nand confer resistance. Based on published results, inhibition of the ADAM17/EGFR ligand axis\nin TNBC should provide therapeutic benefit with reduced tumor growth and decreased\nmetastasis, if not possibly cure.\nIn our in vitro studies on PKC? and PPP1R14D gene knockout in MDA-MB-231 cells indeed led\nto decreases in cellular growth and migration. However, to our surprise, when the same cells\nwere injected into mice through orthotropic fat pad transplantation, they produced aggressive,\nmetastatic tumors that showed activation of alternate growth signaling pathways, namely of the\nmitogen-activated protein kinase ERK and of the PI3kinase target Akt, also a mitogen activated\nkinase. This suggested that TNBC cells were developing resistance to EGF ligand regulator\nknockdown by rewiring their growth factor signaling pathways. To determine where these\nadditional growth signals come from, we first considered the tumor cells themselves.\nIn this context we discovered that when kept in culture, MDA-MB-231 cells expressing sh-\nRNAs targeting either PKCa or PPP1R14D maintained knockdown of the target for up to 35days\ntested. At the same time EGFR and ERK showed low activity as expected due to a decrease in\nEGF ligand cleavage; Akt activity was undetectable. Since we observed strong reactivation of\nERK and new activation of Akt in tumors in vivo, we considered possible up-regulation of other\ngrowth factor receptors on the cell surface that would be engaged by factors released from the\ntumor stroma once cells are inserted in vivo. Indeed, we found that FGFR2 and Erbb4 were\nupregulated. It is therefore likely that reactivation of ERK and new activation of Akt was due to\nFGFR2 and Erbb4. This would suggest that combination therapy of EGF ligand release regulator\ninhibition and FGFR inhibition would decrease growth of these tumors in vivo. Fil: Sharma, Neha. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina Zirlik, Andreas González, Lorena Facultad de Farmacia y Bioquímica Herrlich, Andreas Sharma, Neha 2015-06-01 El cáncer de mama triple negativo (TNCB) es aquel que no expresa el receptor de estrógenos\n(ER), ni el de progesterona (PR) o el HER2. Esta patología representa el 15% de los tumores de\nmama invasivos y tiene una alta incidencia en mujeres jóvenes Afro-Americanas. Es responsable\nde una alta tasa de mortalidad por cáncer de mama ya que generalmente el TNCB causa\nmetástasis; además, responde pobremente a las terapias con quimioterápicos a largo plazo y\ngeneralmente desarrolla resistencia a las terapias dirigidas, incluyendo las que implican al EGFR.\nPor todo ello, es fundamental el desarrollo de terapias alternativas, dado que solo el 30% de las\nmujeres con cáncer de mama metastásico sobrevive pero ninguna de las que presentan TNBC\nmetastásico.\nActualmente, no existe una terapia adecuada y efectiva para el TNBC. En parte, esto se debe a la\nalta heterogeneidad genética que presentan estos tumores, lo cual redunda en la inefectividad de\nterapias basadas en una única droga. Terapias basadas en blancos terapéuticos específicos están\nen investigación y desarrollo, como aquellas basadas en la inhibición de quinasas implicadas en\nseñalización (ejemplo: /Akt, MEK, VEGFR, PDGFR), reparación del DNA, supervivencia\ncelular o acciones androgénicas. Mayormente, estas terapias específicas son combinadas con\nquimioterapia sistémica. Sin embargo, hasta el momento, los beneficios de tales propuestas\nterapéuticas no son claros. Aproximadamente el 60% de los TNBC de tipo basal sobreexpresan\nEGFR; sin embargo, las terapias que implican la inhibición del receptor son mayormente\ninefectivas debido al desarrollo de resistencia. Distintos mecanismos están involucrados en el\ndesarrollo de resistencia a las terapias dirigidas, como ser mutaciones en la proteína blanco o la\nredundancia y sobreactivación de vías de señalización compartidas con otros factores de\ncrecimiento.\nPor lo tanto, es fundamental diseñar terapias combinadas para TNBC que contemplen el posible\ndesarrollo de resistencia. El trabajo de investigación propuesto intenta identificar alteraciones de\nvías de señalización intracelular ocasionadas por las terapias dirigidas, particularmente en lo que\nrespecta al eje ADAM17/EGFR, con el fin de establecer su posible implicancia en el desarrollo\nde resistencia.\nDado que se desconoce como es regulada la actividad y selectividad de ADAM17, se realizó un\namplio estudio mediante shRNA para dilucidar como se regula el clivaje de and PPP1R14D\nregulan el clivaje de TGFa, AREG y HB-EGF sin afectar la actividad proteasa de ADAM17. La\ninhibición del eje ADAM17/EGFR sería beneficioso para el tratamiento del TNBC. Nuestros\nestudios in vitro revelaron que células MDA MB 231 knockout para PKC? and PPP1R14D no\npresentan sobreactivación de RTKs, sugiriendo que en estos modelos podría verse potenciada la\neficacia terapéutica de la inhibición del eje ADAM17/EGFR.\nSin embargo, cuando las mismas células fueron inyectadas a ratones, produjeron un fenotipo de\ntumor agresivo y metastásico, asociado a la reactivación de vías de señalización intracelular\ncomo las mediadas por ERK y PI3K/Akt. Ello se asoció a un aumento de la expresión y\nactivación de distintas RTKs, incluido el EGFR como así también de Akt.\nEstos resultados sugieren la activación alternativa de vías de señalización que permiten que las\ncélulas tumorales proliferen y produzcan metástasis. application/pdf Cocca, Claudia De Siervi, Adriana Mertelsmann, Roland In-Vitro Cáncer Cáncer de mama triple negativo ADAM 17/ EGFR Triple Negative Breast Cancer eng Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/2.5/ar/ Ciencia de la vida Identification of resistance mechanism to targeting of the ADAM 17/ EGFR axis in triple negative breast cancer in vivo info:eu-repo/semantics/masterThesis info:ar-repo/semantics/tesis de maestría info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=afamaster&cl=CL1&d=HWA_835 http://repositoriouba.sisbi.uba.ar/gsdl/collect/afamaster/index/assoc/HWA_835.dir/835.PDF

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