human kidney injury molecule-1 (hkim- 1): a useful immunohistochemical marker for diagnosing renal...
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Journal Reading:Human Kidney Injury Molecule-1 (hKIM-Human Kidney Injury Molecule-1 (hKIM-1): A Useful Immunohistochemical Mark1): A Useful Immunohistochemical Marker for Diagnosing Renal Cell Carcinoma er for Diagnosing Renal Cell Carcinoma and Ovarian Clear Cell Carcinomaand Ovarian Clear Cell Carcinoma Am J Surg Pathol Volume 31, Number 3, March 2007
Fan Lin, MD, PhD,etc
From the Department of Laboratory Medicine, Geisinger Medical Center
Presented by Intern 賴妮均
Introduction (1)
9th most Common malignant tumors in U.S. Notorious for distant metastasis
Lung, lymph node, liver, bone, and brain Subtypes:
Conventional; (clear cell) carcinoma Papillary carcinoma Chromophobe carcinoma
D/D with oncocytoma (benign) Collecting duct carcinoma Unclassified carcinoma
Introduction (2)Immunostaining Markers in RCC
CD10 90% of RCCs
also detected in many hepatocellular carcinomas, prostatic carcinomas, and urothelial carcinomasPositive in normal renal tissue
RCC Ma (RCC marker)
84-92% of primary RCC and 67-84% 0f metastatic RCC
Positive in normal renal tissue
Coexpression of epithelial membrane antigen and vimentin
clear cell RCC
MN/CA9 (Carbonic anhydrase IX)
100% of clear cell RCC and papillary RCC
Also expressed in cervical dysplasia and carcinoma
P504S100% of papillary RCCs and some primary and metastatic clear cell RCC
marker for prostatic adenocarcinomaPositive in normal renal tissue
CD9/MPR1),Antimitochondrialantibody (113–1), E-cadherin and vinculin
high sensitivity for chromophobe RCC, papillary RCC, and oncocytoma
Osteonectin (SPARC) sarcomatoid RCC
Introduction (3)B defensin-1 (+), parvalbumin (+), and vimentin (-)
100 % of Chromophobe RCC and oncocytoma
glutathione S-transferase-a and carbonic anhydrase II(CAII)
Clear cell RCC and chromophobe RCC
kidney-specific cadherin (Ksp-cadherin) andRon proto-oncogene
Majority of both chromophobe RCC and oncocytoma
frequently absent in clear cell RCC Ksp-cadherin :Positive in normal renal tissue
CD117 (c-kit)chromophobe RCC and some oncocytoma
frequently absent in clear cell RCC
S-100 protein70% of clear cell RCC and 90% of oncocytoma
only observed in 6% of chromophobe RCCPositive in normal renal tissue
none of the aforementioned markers is absolutely sensitive and specific for RCC
Introduction (4)
Human kidney injury molecule-1 (hKIM-1)= TIM-1 (T cell immunoglobin domain andmucin domain protein 1)= HAVCR1 (hepatitis A virus cellular receptor 1)
Gene: located in human chromosome 5q Type I transmembranous glycoprotein containing an extracellular immunoglobulinlike domain topping a long mucinl
ike sequence As biomarker for regeneration of renal proximal tubules after tubular injury.3
Material and Method (1)
Case Selection264 cases of renal epithelial neoplasmanonymous and disassociated from any
clinical data
Material and Method (2) TMA sections
221 cases of nonrenal tumors from various organs to test the specificity of anti-hKIM-1 antibody
Material and Method (3)
TMA constructionH&E stain sections were reviewedremove 1.0-1.5 mm tissue core from marked
area of paraffin block inserted into recipient paraffin block
(Beecher Instruments Inc, Sun Prairie, WI)
Material and Method (4)
Immunohistochemistry Formalin-fixed and paraffin-embedded 4μm sections Antibody to hKIM-1
AKG7 monoclonal antibody Confirmed by ABE3 antibody in CCCO and uterus
Positive control: Kidney tissues with acute tubular necrosis
Negative control: replacement of the primary antibody with nonimmune mouse serum
Interpretation: 2 surgical pathologists indicates negative; 1+, <10%; 2+, 11% to 50%; 3+, >50%.
Material and Method (5)
Western Blot 4 cases of renal tumor with different subtypes Normal renal tissue from a clear cell RCC case
Frozen tissue blocksLysed tissue homogenatws Electrophoresed on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) Transferred onto polyvinyl diflouride membranes Incubated with AKG7 monoclonal antibody
Evenness of the protein loading: antibody to β-actin
Material and Method (6)
Gene Expression Microarray tissues were frozen promptly after surgery an
d stored at -80 .℃hKIM-1 mRNA level
Results (1) Non-neoplastic renal tissue
Negative: Glomeruli, vessels, inflammatory cells, interstitialcells, lymphoid aggregatesinterstitial tissues
Positive immunoreactivity was observed in proximal tubules, with a cytoplasmic / membranous staining pattern during acute tubular necrosis
2 of 20 case in TMA showed focal immunostaining occasional proximal tubules
Result (2)
Renal Epithelial Neoplasm
(2/15) in TMA
Result (3)Clear cell RCC, Papillary RCC
Result (4)
Chromophobe RCC
Result (5) Diffuse immunoreactivity was more frequently observed in high-grad
e clear cell RCC (56%) than in low-grade clear cell
2/2 sacromatoid RCC were positive 1 case of Wilms tumor and no immunoreactivity was observed (data not shown)
Result (6)
Metastatic clear cell RCC The percentages of positive c
ases were not significantly different among theseanatomic sites.
Result (7) Non-renal Neoplasms
Result (8) CCCO
Result (9) Clear cell carcinoma of uterus Adenocarcinoma of the colon
Result (10)
Western Blot
Result (11) hKIM-1 mRNA in Renal Tumors From Affy
metrix Microarrays
0
-131
537
285
-126
Discussion
Do we really need another immunohistochemical marker?
Discussion
Differentiated from normal renal tissue The previously reported biomarkers are expressed in normal renal tissue Murine KIM-1 expressed in actived T cells and linked to air hypersensitivity, but h
KIM-1 does not
Negative staining in: Non-neoplastic renal tissues Transplanted renal biopsy with acute cellular rejection
Positive staining in: (1) proximal tubules adjacent to renal tumors (2) proximal and distal tubules in the tumorous kidney, probably secondary to tumor ob
struction and compression (3) atrophic renal tubules; (4) proximal tubules associated with tubular injury,such as hypoxia, ischemia, toxin, inf
ection, etc
Discussion
More specific to clear cell RCC and papillary RCC Oncocytoma: only 4 of 41 (9.8%) with focally positive stained Chromophobe RCC: all negative
CD-10 also expressed in oncocytoma RCC Ma also expressed in chromophobe RCC P504S also expressed in both oncocytoma and chromophobe
RCC S-100 also expressed in >90% oncocytoma
Discussion
More specific to metastatic RCC Han et al: only 1 of 450 nonrenal tumor was positive
(HCC) CD10, RCC Ma, S-100, P504S, glutathione S-transfer
ase-a, CD117: well-documented in other nonrenal tumors
Why higher positive reaction than previoud Han’s study?
CCCO and uterus were not included The previous study only checked TMA
Discussion
What are the unique features and utility of this antibody?
Discussion
Expression of hKIM-1 in CCCO (93.8%) Both ovary and uterus serous carcinoma and endometroid carcinoma la
cked hKIM-1 expression Dent and coworkers: gene amplification in CCCO was most common at
chromosome 3 Von Hippel-Lindau tumor-suppressor genes:
located at the chromosome 3p25; and the von Hippel-Lindau tumorsuppressor
closely linked to the tumorigenesis of both sporadic and hereditary clear cell RCC
hKIM-1 had similar role in both clear cell RCC and CCCO? Anti-hKIM-1Ab may be used in targeted cancer therapy for CCCO?
Discussion
clear cell carcinoma of the uterusonly 33% expressed hKIM-1not related to chromosome 3 alternation
Clear cell carcinoma of uterus Adenocarcinoma of the colon
Discussion
Focal positive staining in adenocarcinoma of the colon 5 of 40 cases Pattern: apical portions of neoplastic glands
Only in well and moderately differentiatied colonic adenocarcinoma
the AKG7 anti-hKIM-1 monoclonal antibody:against the extracellular mucin domaincross reaction
Disscusion
Conventional RCC S-100+/hKIM-1+/MSH2-/CK7-
Chromophobe RCC MSH2+ (cytoplasmic staining) /CK7+/S-100-/hKIM-1-
Oncocytoma S-100+/MSH2+ (apical and nuclear staining) /hKIM-1-/CK7-or fo
cal +
Conclusion
HKIM-1 is a relatively sensitive and specific marker for confirming the diagnosis of papillary RCC, cle
ar cell RCC, and metastatic RCCdistinguishing clear cell RCC from chromopho
be RCC and oncocytoma identifying CCCO
Thanks for your attention !