cancer dx algorithm
DESCRIPTION
This is needs to be updated as new Biomarkers and Dx antibodies come into existence in remarkable numbers every day! You have to construct an algorithm based on your need and availability of antibodies. Selection of Antibodies for the algorithm also depends on your ability to recognize the characterization of the specific antibody.TRANSCRIPT
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Mission: Mission: We connect to fight cancer!We connect to fight cancer!
As a world-leading expert DakoAs a world-leading expert Dako continuously challenges and raises the continuously challenges and raises the global standards for fast and accurate global standards for fast and accurate answers for cancer patients and doctors answers for cancer patients and doctors making vital decisions for treatmentmaking vital decisions for treatment
CONNECTING REAGENTS,INSTRUMENTS AND
SOFTWARE
Shaping the future of cancer diagnosis
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Diagnosis of Diagnosis of CancerCancer using using
AlgorithmsAlgorithms
Lawrence T. RichardsLawrence T. RichardsM.S.,H.T(ASCP).,QIHC(ASCP).M.S.,H.T(ASCP).,QIHC(ASCP).
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This presentation is by no means intended as a replacement of the professional judgment of a certified pathologist. The contents of the algorithm are provided as informative only, and should not be regarded as indicative of standard procedure for diagnosis or treatment.
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Diagnosis of Cancer using Diagnosis of Cancer using AlgorithmsAlgorithms
A diagnostic algorithm is a method which A diagnostic algorithm is a method which utilizes a panel of antibodies intended to utilizes a panel of antibodies intended to solve a diagnostic problemsolve a diagnostic problem
Many different diagnostic algorithms exist Many different diagnostic algorithms exist and are available in journals and text and are available in journals and text booksbooks
A diagnostic algorithm is followed by a A diagnostic algorithm is followed by a selective markers for tumor sub selective markers for tumor sub classification.classification.
The panel of antibodies selected should be The panel of antibodies selected should be based on the morphological appearance of based on the morphological appearance of the tissue (H & E) and the patient’s the tissue (H & E) and the patient’s clinical history provided by the physician.clinical history provided by the physician.
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MUC-1 PAS
MUC-1(IHC) PAS (Special Stain)
AdenoCarcinoma
Special stain vs IHCSpecial stain vs IHC
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Markers and HistogenesisMarkers and Histogenesis HISTOGENESISHISTOGENESIS MARKERSMARKERS MesenchymalMesenchymal VimentinVimentin EpithelialEpithelial Cytokeratin, EMACytokeratin, EMA Smooth MuscleSmooth Muscle Desmin, HHF35, Desmin, HHF35,
SmActinSmActin Skeletal MuscleSkeletal Muscle MyoglobinMyoglobin FibrohistiocyteFibrohistiocyte CD68, Factor XIIIaCD68, Factor XIIIa Nerve SheathNerve Sheath Leu7,Leu7,Glial fibrillary acidic Glial fibrillary acidic
proteinprotein MelanocyteMelanocyte HMB 45HMB 45 Neuronal Neuronal Neurofilament Neurofilament Endothelial, perivascularEndothelial, perivascular Factor Factor
VIII,CD34,CD31VIII,CD34,CD31 HematopoiticHematopoitic LCA,CD3,CD20,Ki-1LCA,CD3,CD20,Ki-1 NeuroendocrineNeuroendocrine
NSE,Chromogranin,SynaptophysinNSE,Chromogranin,Synaptophysin Ewing’s sarcoma/PNETEwing’s sarcoma/PNET MIC-2(O-13)MIC-2(O-13)
Source:www.moffitt.usf.edu/pubs/ccj/v5nl/department5/htmlSource:www.moffitt.usf.edu/pubs/ccj/v5nl/department5/html
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Basic Flow ChartBasic Flow ChartSequentialSequential
First Round
(+) ( - )
(+)
(+)
(+)
( - )
( - )
( - )
Second round
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Use of Panel of Use of Panel of AntibodiesAntibodies
Ab1 Ab1 Ab2Ab2 Ab3Ab3 Ab4Ab4 Ab5 Ab5 Ab6Ab6
+ (-)+ (-) + + (-)(-) + + + Tumor-1+ Tumor-1
(-)(-) ++ (-)(-) ++ (-)(-) (-) Tumor-2(-) Tumor-2
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Differential ApplicationDifferential Application
Tumor 1Tumor 1 Tumor 2Tumor 2 Ab1 +Ab1 + Ab1 –Ab1 – Ab2 -Ab2 - Ab2 +Ab2 +
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Diagnostic AlgorithmDiagnostic Algorithmfor unknown Tumor Typefor unknown Tumor Type
CarcinoCarcinomama
LymphoLymphomama
MelanoMelanomama
SarcomaSarcoma NotesNotes
Pan Pan KeratinKeratin
PositivPositivee
NegativeNegative NegativeNegative NegativeNegative ForFor
Poorly Poorly differentiated differentiated malignant malignant tumor tumor differentiationdifferentiation
Caution:Caution:
Variation in Variation in cellular cellular pattern can pattern can misleadmislead
(+)(+) and and (-)(-)..
LCALCA NegativeNegative PositivPositivee
NegativeNegative NegativeNegative
S 100 / S 100 / HMB45/HMB45/
MART-1MART-1
TyrosinaseTyrosinase
NegativeNegative NegativeNegative PositivPositivee
NegativeNegative
DesminDesmin NegativeNegative NegativeNegative NegativeNegative PositivPositivee
VimentinVimentin NegativeNegative NegativeNegative PositivePositive PositivPositivee
Source adopted from: DAKO training manual
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Undifferentiated Undifferentiated Malignant NeoplasmMalignant Neoplasm
Abs toAbs to Abs/Abs/cloneclone
CarcinoCarcinomama
LymphoLymphomama
MelanoMelanomama
SarcomaSarcoma
CytokerCytokeratinatin
AE1/AE3AE1/AE3
VimentinVimentin V9V9
MelanoMelanoma ma antigenantigen
HMB 45HMB 45
CD 45CD 45 PD7/2B1PD7/2B111
CD43CD43
Almost always positive
Almost always Negative Sometimes positive
Sometimes negative
Gowen AM.Diagnostic Immunohistochemistry of Solid Tumors: Strategies and Solutions,USCAP;2002 #35
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Carcinoma of Carcinoma of Unknown PrimaryUnknown Primary
The hypothesis is that the The hypothesis is that the primary tumor either remains primary tumor either remains
microscopic and escapes clinical microscopic and escapes clinical detection detection oror disappears after disappears after
seeding the metastasis.seeding the metastasis.
Antibody AlgorithmAntibody Algorithm is used to is used toSearch for primary SiteSearch for primary SiteRule out Non-Carcinoma Rule out Non-Carcinoma
Like lymphoma, melanoma, Like lymphoma, melanoma, sarcomasarcoma
Identify Sub-Groups for Identify Sub-Groups for TreatmentTreatment
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Undifferentiated panel: Ca vs Mel vs La Keratin AE1/3 mix, 90%+ of Ca Not CK7 or 20 or 5/6 to start with EMA, CEA as backups S100, 95%+ of Mel 10% of Ca are + HMB45, MelanA less sensitive, very specific LCA or CD20 90% of lymphomas Things that might be neg Anaplastic large cell lymphoma - use CD30, ALK Plasmacytoma – use CD138, kappa, lambda Sarcomas – use various markers, esp vascular Spindled/sarcomatoid carcinoma – use CK5/6 and p63 Liver – use HepPar1 Adrenal – use Inhibin, MelanA Seminoma/germinoma – use OCT3/4
http://surgpathcriteria.stanford.edu/IPOXHandout.pdf
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: >90% of tumors positive*
: 50-90% of tumors positive*
:
10-50% of tumors positive*
: <10% of tumors positive*
? : Insufficient evidence[ ]
: Staining of secondary (non-neoplastic) cell type
( )
:Staining dependant on antibody
* Positive Generally : unequivocal staining of =10% of tumor cells
Threshold for PositivityThreshold for Positivity
+/-
+
-/+
(-)
Source: Dako antibody algorithm
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Metastatic Adenocarcinoma ofMetastatic Adenocarcinoma of unknown origin
with site specific markerswith site specific markers
PSA+(-)
(-)
(-)
(-)
(-)
(-)
(-)
(-)
+
+
+
+
+
+
+
Prostate
Lung
Stomach/PancreasBreast
Colon
Colon
Stomach / Pancreas
Breast
Ovary
Pancreas,(Ovary serous)
Stomach / Pancreas
Breast / Stomach / Pancreas
Source: www.clincancerres.aacrjournals.org/egi/content/full/11/10/3766
TTF-1
GCDFP15
CDX2 / CK20
ER
CA125
Mesothelin
Lysozyme
+
+
+
(-)
(-)
(-)
CDX2
CK 7
Mesothelin
MC5+ (98%)
PE10+TTF-1=100%Lung
CK8+CK18=100% AdCa
CK7(+)
All (-) do HEPPAR-1 for HCC
Mixed pattern
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Differential DiagnosisDifferential Diagnosis Bladder vs Bladder vs
Prostate ca Prostate ca
Cytokeratin Cytokeratin 77
++ (-)(-)
Cytokeratin Cytokeratin 2020
++ (-)(-)
CEACEA ++ (-)(-)
PAPPAP (-)(-) ++
PSAPSA (-)(-) ++
Goldstein N.Immunohistochemical Antibody Panels to help Identify the Primary Sites of Various Carcinomas;ASCP Teleconference # 2109
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Mesothelioma or
Small Cell Carcinoma ?
Mesothelioma=BG8(-);Calretinin(+);MOC-31(-)
SmallCellCa= TTF-1 (+);Leu-7(+)
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Mesothelioma vs CarcinomaMesothelioma vs CarcinomaAntigenAntigen Mesothelioma %Mesothelioma % Carcinoma %Carcinoma %
CytokeratinCytokeratin 100100 100100
EMAEMA 8080 100100
ThrombomodulinThrombomodulin 100100 88
Leu-M1Leu-M1 1010 7575
S-100S-100 1010 8585
CEACEA 00 1515Placental Alkaline Placental Alkaline PhosphatasePhosphatase
00 6565
VimentinVimentin 4040 3030
Ber-EP4Ber-EP4 00 8080
CalretininCalretinin 9090 3030
B72.3B72.3 00 7070Brown RW et al.Multiple-marker immunohistochemical phenotypes distinguishing malignant Brown RW et al.Multiple-marker immunohistochemical phenotypes distinguishing malignant pleural mesothelioma from pulmonary adenocarcinoma. Hum Pathol.1993;24:347-354pleural mesothelioma from pulmonary adenocarcinoma. Hum Pathol.1993;24:347-354
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Cytokeratin Cytokeratin phenotypephenotype CK phenotypeCK phenotype CK7 (-) / CK 20 CK7 (-) / CK 20
(-)(-)
CK7 (+) / CK 20 CK7 (+) / CK 20 (-)(-)
CK7 (-) / CK 20 CK7 (-) / CK 20 (+)(+)
CK7 (+) / CK 20 CK7 (+) / CK 20 (+)(+)
TumorsTumors HNC, Liver, Lung (SqCC and HNC, Liver, Lung (SqCC and
SmCC),Prostate, RenalSmCC),Prostate, Renal
Biliary and Pancreas, Breast, Biliary and Pancreas, Breast, Cervical, EM, Lung(Ad ca), Cervical, EM, Lung(Ad ca), Ovarian (non-mucinous), Ovarian (non-mucinous), thyroidthyroid
Colon, Gastric, Markel C caColon, Gastric, Markel C ca
Biliary and Pancreas, Ovarian Biliary and Pancreas, Ovarian (Mucinous), Urothelial.(Mucinous), Urothelial.
Source: from Adopted Dennis JL, Clin Cancer Res 2005
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Cytokeratin for Carcinoma Cytokeratin for Carcinoma DiagnosisDiagnosis
CK5CK5 CK7CK7 CK20CK20Breast Ca DuctalBreast Ca Ductal ★★ ● ● ○ ○Lung nonsmall CellLung nonsmall Cell ○ ○ ● ● ○ ○ (non SqCCa) (non SqCCa) Lung SqCCaLung SqCCa ● ● ★ ★ ○ ○Pancreatic Ca Pancreatic Ca ○ ○ ★ ★ ★ ★ Colorectal Ad Ca Colorectal Ad Ca ○ ○ ○ ○ ● ●Mesothelioma Mesothelioma ● ● ● ● ○ ○Prostatic Ad CaProstatic Ad Ca ○ ○ ○ ○ ○ ○ Transitional Cell CaTransitional Cell Ca ● ● ● ● ● ●
★ ★ May be Positive ● Positive ○ NegativeMay be Positive ● Positive ○ Negative
Source: A.M.Gown ASCP Course #35Source: A.M.Gown ASCP Course #35
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Prostate Ca or Prostate Ca or Benign ?Benign ?
Prostate Prostate CancerCancer EpCam +EpCam + ATM +ATM + AMACR +AMACR + PSA + / (-)PSA + / (-) 3434ßE12 (-) almost allßE12 (-) almost all p63 (-) almost allp63 (-) almost all Prostein +Prostein + NKX3.1 + NKX3.1 +
Benign Benign ProstateProstate
EpCam (-)EpCam (-) ATM (-) / +ATM (-) / + AMACR (-)AMACR (-) PSA + / (-)PSA + / (-) 3434ßE12 +ßE12 + p63 +p63 + Prostein +Prostein + NKX3.1 + NKX3.1 +
Hammerich KH et al.Application of Immunohistochemistry to the Genitourinary System(Prostate, Urinary Bladder, Testis, and Kidney)Archives of Pathology and Laboratory Medicine;132:432-440,2007
ATM=ataxia-telangiectasia mutated;AMACR=alpha-methylacyl-CoA racemase;Ep-Cam=epithelial transmembrane glycoprotein
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Urothelial Ca vs Urothelial Ca vs Prostate CaProstate Ca
EMAEMA CK7CK7 P63P63 CK5/6CK5/6 EpCamEpCam CD57CD57 PSAPSA PAPPAP NKX3.NKX3.
11 ProsteiProstei
nn
++ --++ --+/-+/- 00+/-+/- 00+/-+/- 00
-/+-/+ +/-+/- 00 ++
00 +/-+/- 00 ++ 00 ++ 00 ++
Urothelial ca Prostate ca
Hammerich KH.Archives of Pathology and Laboratory Medicine;132(3):432-440
Urothelial ca
Prostate ca
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Ab CocktailsAb Cocktails Breast CaBreast Ca: : ER + GCDFP-15ER + GCDFP-15 MelanomaMelanoma: : MART-1+Melan MART-1+Melan
A+Tyrosine +A+Tyrosine +
PNL2PNL2 Basal Cell MarkerBasal Cell Marker: : p63 + 34betaE12 p63 + 34betaE12
+ +
CK5/6CK5/6 Endocrine Ca:Endocrine Ca: Chrom A+NSE +Chrom A+NSE +
SynaptophysinSynaptophysin Pan SarcomaPan Sarcoma: Vim + Coll IV + CD 99: Vim + Coll IV + CD 99
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Endocervical Ad.Ca & Endocervical Ad.Ca & Endometrial Mucinous Endometrial Mucinous
Ad.CaAd.CaECAECA MUC-1(-)MUC-1(-) ER(-)ER(-) PR(-)PR(-) P16(+)P16(+)
EMMAEMMA MUC-1 (+)MUC-1 (+) ER(+)ER(+) PR(+)PR(+) P16(-)P16(-)
Khoury T et al.BMC Clin Path 2006;6:1
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A, Normal post-menopausal endometrium (H&E, 20x); B, MUC1 immunostain in normal postmenopausal endometrium showing “pure apical” staining pattern (x20); C, normal endocervical glands (H&E, 20x); D, MUC1 immunostain in normal endocervical glands showing A/C staining pattern, (x20).
MUC1-Purely Apical vs Apical & Cytoplasmic
Khoury T, et al.BMC Clin Pathol.2006;6:1
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A, Endocervical adenocarcinoma (H&E, x20); B, MUC1 immunostain in endocervical adenocarcinoma showing apical staining pattern (x20).
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A, Endometrial mucinous adenocarcinoma (H&E, x10); B, MUC1 immunostain in endometrial mucinous adenocarcinoma showing A/C staining pattern (x10); C, MUC1 immunostain in endometrial mucinous adenocarcinoma showing A/C staining pattern (x40).
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The Cost-Effectiveness of The Cost-Effectiveness of IHCIHC
Why do IHC assay?Why do IHC assay? To increase patient life expectancy by correctTo increase patient life expectancy by correct
diagnosis and treatment plan.diagnosis and treatment plan. To increase diagnostic certaintyTo increase diagnostic certainty To predict patient prognosisTo predict patient prognosis
eg., 35yr old female, history of Br ca, radical eg., 35yr old female, history of Br ca, radical mastectomy performed earlier. Now mastectomy performed earlier. Now diagnosed with cirrhosis and liver mass.diagnosed with cirrhosis and liver mass.
1.1. Meatastatic Ad Ca ?Meatastatic Ad Ca ?2.2. Hepato Cellular Ca ?Hepato Cellular Ca ?
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http://www.dako.com/asp/algo/http://www.dako.com/asp/algo/default.asp?default.asp?
get=disclaimer&type=noget=disclaimer&type=noContact:
Gitte SjorslevMail: [email protected]
DAKO Cancer Diagnostic Algorithms
DAKO Cancer Diagnostic Algorithms
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Navigating DAKO Cancer Navigating DAKO Cancer Diagnostic Algorithm.Diagnostic Algorithm.
Terms and Conditions for use - Dako Antibody AlgorithmTerms and Conditions for use - Dako Antibody Algorithm
Antibody Algorithm - Contents Antibody Algorithm - Search eg.,
ChooseSearchTumor Antibody
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Table 1
Table 1
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Table 2.0
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Table 2.1
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LastlyLastly
“ “The Diagnostic Power of any The Diagnostic Power of any Immunohistochemical Procedure Immunohistochemical Procedure is no Greater than the wisdom of is no Greater than the wisdom of the Pathologist interpreting it.” the Pathologist interpreting it.”
Dr.Allen GownDr.Allen Gown
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http://www.ascp.org/pdf/5604TableofContents.aspx
Efficient Tumor ImmunohistochemistryA Differential Diagnosis-Driven Approach
M. Nadji, MD, M. Nassiri, MD,and A.R. Morales, MD
http://www.nordiqc.com/
http://appliedimmuno.org/
http://surgpathcriteria.stanford.edu/IPOXHandout.pdf
http://www.immunoportal.com/index.php
http://web.ncifcrf.gov/rtp/lasp/phl/immuno/
http://www.phenopath.com/diagnostic/teaching.htm
Web Sites
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Thanks to
DAKO
&
MMJ Biosystems Philippines, Inc.
Address: Unit 206 CYA Land Bldg. (formerly LTC Corporate Center Bldg.)
#282 EDSA Extension Cor. P. Celle Street, Pasay City, Manila
Tel. nos.: (632) 851-0192 to 93/489-1008Telefax: (632) 853-3665
email: [email protected]
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38 www.markwickmd.com/documents/UNDIFF_TMRS2.ppt
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www.biomedcentral.com/1472-6890/6/1/figure/F3?highres=ywww.moffitt.usf.edu/pubs/ccj/v5nl/department5/htmlGowen AM.Diagnostic Immunohistochemistry of Solid Tumors: Strategies and Solutions,USCAP;2002 #35Source: www.clincancerres.aacrjournals.org/egi/content/full/11/10/3766Goldstein N.Immunohistochemical Antibody Panels to help Identify the Primary Sites of Various Carcinomas;ASCP Teleconference Brown RW et al.Multiple-marker immunohistochemical phenotypes distinguishing malignant pleural mesothelioma from pulmonary adenocarcinoma. Hum Pathol.1993;24:347-354Dennis JL, Clin Cancer Res 2005Source: A.M.Gown ASCP Course #35Hammerich KH et al.Application of Immunohistochemistry to the Genitourinary System(Prostate, Urinary Bladder, Testis, and Kidney)Archives of Pathology and Laboratory Medicine;132:432-440,2007Hammerich KH.Archives of Pathology and Laboratory Medicine;132(3):432-440Khoury T et al.BMC Clin Path 2006;6:1www.markwickmd.com/documents/UNDIFF_TMRS2.ppt
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ER by Nadji M
Appl Immunohistochem Mol Morphol. 2008 Mar;16(2):105-7
“Seasoned immunohistochemists, nevertheless, know that IHC of routinely fixed and processed tissue does not yield itself to accurate quantitation of results, even when performed by well-qualified laboratories. Furthermore, in the case of ER, immunohistochemical methods only identify a segment or epitope of ER protein that is immunologically reactive with the used antibody. Hence, as it is, an immunohistochemical technique gives no information about the functional status of ER molecule, and/or that of the complex downstream ER pathways. This may be one of the reasons why one-third of patients with ER-positive breast cancers initially, and another one-third eventually, do not respond to endocrine treatment modalities. In this review, I attempt to present an argument that is based on our current information; quantitation of ER-IHC is neither”