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Identification of tissue biomarkers of prognostic significance in pancreatic cancer

Hu, Dingyuan LU (2019)
Abstract (Swedish)
Pankreascancer, dvs cancer i bukspottkörteln, utgör nu den tredje vanligaste orsaken till död i cancer och kommer om inga genombrott görs att utgöra andra orsak till cancerrelaterad död inom några år (efter lungcancer). Avsaknad av tidiga symtom och en uttalad terapisvikt är bidragande faktorer till den dåliga prognosen. Femårsöverlevnaden ligger på endast maximalt 6 % och pankreascancer orsakar, förutom den höga dödligheten, också betydande kostnader för samhället.
Det finns idag ett mycket begränsat antal diagnostiska biomarkörer för såväl blodprov som vävnadsdiagnostik. I serum har sedan decennier tumörmarkören CA 19-9 använts medan man i vävnad inte haft några möjligheter att identifiera biomarkörer som skulle ge en mera... (More)
Pankreascancer, dvs cancer i bukspottkörteln, utgör nu den tredje vanligaste orsaken till död i cancer och kommer om inga genombrott görs att utgöra andra orsak till cancerrelaterad död inom några år (efter lungcancer). Avsaknad av tidiga symtom och en uttalad terapisvikt är bidragande faktorer till den dåliga prognosen. Femårsöverlevnaden ligger på endast maximalt 6 % och pankreascancer orsakar, förutom den höga dödligheten, också betydande kostnader för samhället.
Det finns idag ett mycket begränsat antal diagnostiska biomarkörer för såväl blodprov som vävnadsdiagnostik. I serum har sedan decennier tumörmarkören CA 19-9 använts medan man i vävnad inte haft några möjligheter att identifiera biomarkörer som skulle ge en mera individbaserad behandling och därmed potentiellt kunna öka överlevnad.
Syftet var att identifiera och validera nya biomarkörer från pankreascancervävnad för en förbättrad prognostisk information.
Med hjälp av s k proteomik och en teknik benämnd masspektrometri studerades initialt formalin-fixerad paraffininbäddad cancervävnad från nio patienter som genomgått pankreaskirurgi med en efterföljande kort överlevnad (< 12 månader), respektive tio patienter med en förhållandevis lång överlevnad (> 45 månader). Slutligen definierades 25 proteiner, varav 18 var uppreglerade vid lång överlevnad och sju uppreglerade vid kort överlevnad.
I de åtföljande fyra delarbetena utförs ett valideringsarbete på cirka 140 patienter som genomgått kirurgisk resektion av sin pankreascancer. Teknikerna som genomgående använts var konstruktion av en s k tissue microarray, dvs ett mycket litet vävnadsblock, som sedan snittas och infärgas med immunhistokemi riktat mot de olika markörer som identifierats i den initiala proteomikstudien.
I delarbete 2 studeras calcium activate chloride channel regulator-1 (CLCA1) där en låg nivå visar sig korrelera med en kortare sjukdomsfri överlevnad.
I delarbete 3 befinnes markören galectin-4 korrelera till återfall av pankreascancer inom ett år och korrelerar också i sin nivå med den totala överlevnaden både ett och tre år efter kirurgi.
I delarbete 4 kommer vi in på de betydelsefulla fynd man har vid pankreascancer, dvs att enbart 10-20 % av tumören utgörs av cancerceller medan kringliggande bindväv med såväl specifika celler och kollagen dominerar och har en koppling till tumörens tillväxt och metastaseringspotential.
I det fjärde arbetet visade sig lågt uttryck av P4HA2, som bildar kollagen, och ett högt uttryck av PRTN3, som bryter ner kollagen, där denna kombination korrelerar till en mycket kort både sjukdomsfri som total överlevnad.
I det femte delarbetet studeras fibronectin-1-uttrycket i stroma där ett högt uttryck korrelerar med aggressiva tumörkaraktäristika på opererade patienter, men någon koppling till överlevnad kan dock inte ses.
Sammanfattningsvis finns här nu en “panel” med nya pankreascancerbaserade biomarkörer som ger prediktiv och prognostisk information. Ytterligare valideringsarbete kommer att göras. (Less)
Abstract
Background: Pancreatic cancer is the third leading cause of cancer-related mortality. Lack of early detection strategies and therapeutic resistance are main contributors to the poor prognosis. Unfortunately, there are no tissue biomarkers available for the prognosis of pancreatic cancer in routine clinical use.Aim: To identify and validate novel tissue biomarkers for the prognosis of pancreatic cancer.Methods: A mass spectrometry-based proteomic approach was applied to formalin-fixed paraffin-embedded specimens from surgically resected pancreatic cancer in 9 patients with short survival (<12 months) and 10 patients with long survival (>45 months). The dysregulated biomarkers were further verified by targeted proteomics, parallel... (More)
Background: Pancreatic cancer is the third leading cause of cancer-related mortality. Lack of early detection strategies and therapeutic resistance are main contributors to the poor prognosis. Unfortunately, there are no tissue biomarkers available for the prognosis of pancreatic cancer in routine clinical use.Aim: To identify and validate novel tissue biomarkers for the prognosis of pancreatic cancer.Methods: A mass spectrometry-based proteomic approach was applied to formalin-fixed paraffin-embedded specimens from surgically resected pancreatic cancer in 9 patients with short survival (<12 months) and 10 patients with long survival (>45 months). The dysregulated biomarkers were further verified by targeted proteomics, parallel reaction monitoring. Finally, we evaluated prognostic candidates (CLCA1, galectin 4, P4HA2, PRTN3 and fibronectin) by tissue microarray and immunohistochemistry in a larger cohort of patients with pancreatic cancer who underwent surgical resection (n=144). Bioinformatic analysis was exploited to assess pathways and networks linked to the prognosis. Kaplan-Meier and Cox proportional hazards modeling were used to explore the association between biomarkers and survival.Results/Conclusion: A total of 24 and 147 proteins were significantly upregulated in patients with short survival and long survival, respectively. Bioinformatic analysis linked proteins representing “activated stroma factors” and “basal tumor factors” to poor prognosis and highlighted TCF1 and CTNNB1 as possible upstream regulators. By targeted proteomics, seven proteins were verified to be upregulated in patients with short survival (MMP9, CLIC3, MMP8, PRTN3, P4HA2, THBS1 and FN1), while 18 proteins were upregulated in patients with long survival, including EPCAM, galectin 4, VIL1, CLCA1 and TPPP3 (I). By immunohistochemical validation, we found that low CLCA1 expression correlated significantly with shorter disease-free survival (II). Furthermore, galectin 4 expression significantly correlated with disease recurrence within 1 year of surgery and with overall survival at 1- and 3-year (III). Besides, a low P4HA2 and high PRTN3 expression pattern correlated with shorter disease-free survival and overall survival (IV). Finally, high stromal FN1 expression was associated with aggressive tumor characteristics in patients with resected pancreatic cancer, although it was not associated with survival (V). (Less)
Please use this url to cite or link to this publication:
author
supervisor
opponent
  • professor Sund, Malin, Umeå University, Umeå, Sverige
organization
alternative title
Identifiering av vävnadsbiomarkörer av prognostisk betydelse vid pankreascancer
publishing date
type
Thesis
publication status
published
subject
keywords
pancreatic cancer, proteomics, mass spectrometry, Biomarkers, prognosis, CLCA1, pankreascancer, proteomik, masspektrometri, biomarkörer, prognos, CLCA1
pages
102 pages
publisher
Lund University, Faculty of Medicine
defense location
Föreläsningssal 1, Centralblocket, Entrégatan 7, Skånes Universitetssjukhus i Lund
defense date
2019-05-13 13:00
ISBN
978-91-7619-777-6
language
English
LU publication?
yes
id
5a1825b0-df0c-4e1b-ba7d-875a8e911044
date added to LUP
2019-04-12 18:07:37
date last changed
2019-04-15 15:31:01
@phdthesis{5a1825b0-df0c-4e1b-ba7d-875a8e911044,
  abstract     = {Background: Pancreatic cancer is the third leading cause of cancer-related mortality. Lack of early detection strategies and therapeutic resistance are main contributors to the poor prognosis. Unfortunately, there are no tissue biomarkers available for the prognosis of pancreatic cancer in routine clinical use.Aim: To identify and validate novel tissue biomarkers for the prognosis of pancreatic cancer.Methods: A mass spectrometry-based proteomic approach was applied to formalin-fixed paraffin-embedded specimens from surgically resected pancreatic cancer in 9 patients with short survival (&lt;12 months) and 10 patients with long survival (&gt;45 months). The dysregulated biomarkers were further verified by targeted proteomics, parallel reaction monitoring. Finally, we evaluated prognostic candidates (CLCA1, galectin 4, P4HA2, PRTN3 and fibronectin) by tissue microarray and immunohistochemistry in a larger cohort of patients with pancreatic cancer who underwent surgical resection (n=144). Bioinformatic analysis was exploited to assess pathways and networks linked to the prognosis. Kaplan-Meier and Cox proportional hazards modeling were used to explore the association between biomarkers and survival.Results/Conclusion: A total of 24 and 147 proteins were significantly upregulated in patients with short survival and long survival, respectively. Bioinformatic analysis linked proteins representing “activated stroma factors” and “basal tumor factors” to poor prognosis and highlighted TCF1 and CTNNB1 as possible upstream regulators. By targeted proteomics, seven proteins were verified to be upregulated in patients with short survival (MMP9, CLIC3, MMP8, PRTN3, P4HA2, THBS1 and FN1), while 18 proteins were upregulated in patients with long survival, including EPCAM, galectin 4, VIL1, CLCA1 and TPPP3 (I). By immunohistochemical validation, we found that low CLCA1 expression correlated significantly with shorter disease-free survival (II). Furthermore, galectin 4 expression significantly correlated with disease recurrence within 1 year of surgery and with overall survival at 1- and 3-year (III). Besides, a low P4HA2 and high PRTN3 expression pattern correlated with shorter disease-free survival and overall survival (IV). Finally, high stromal FN1 expression was associated with aggressive tumor characteristics in patients with resected pancreatic cancer, although it was not associated with survival (V).},
  author       = {Hu, Dingyuan},
  isbn         = {978-91-7619-777-6},
  keyword      = {pancreatic cancer,proteomics,mass spectrometry,Biomarkers,prognosis,CLCA1,pankreascancer,proteomik,masspektrometri,biomarkörer,prognos,CLCA1},
  language     = {eng},
  pages        = {102},
  publisher    = {Lund University, Faculty of Medicine},
  school       = {Lund University},
  title        = {Identification of tissue biomarkers of prognostic significance in pancreatic cancer},
  year         = {2019},
}