JAMA. Patients with other than HNSCC such as adenocarcinoma, melanoma, sarcoma and metastasis were excluded. Two studies 25, 26 provided results for p16 INK4a interpretation as p16 INK4a positive or negative, . The results discussed above demonstrate that when p16 IHC is utilized based on the LAST criteria there is an improvement in the overall performance of ISPs compared with the CPR. However, the LAST criteria were developed specifically with a desire to discourage and prevent inappropriate use or overuse of any biomarker(s).12 As a result, they may be overly restrictive as to which biopsies would benefit from p16 IHC. These differences could also be attributed to the difference in geographic distribution of tumor, sample size, scoring criteria and different types of antibodies used by different authors. As ISPs had read all cases without and with p16 in the 2 reading rounds in the course of the study, it was possible to assess the effect of adjunctive p16 slide interpretation also within the cases for which the ISPs did not request p16 during their first reading round on H&E-stained slides (designated as non-LAST cases in this analysis). 2021 Oct 5;14:1419-1425. doi: 10.2147/CCID.S333376. The correlation of p16 expression with lymphadenopathy was found to be highly statistically significant (P = 0.009). It involved 1100 cervical biopsies and 70 ISPs, each of whom read the biopsies twice, once using only H&E-stained slides (ISPH&E) and a second time using both H&E and p16-stained slides (ISPH&E+p16). Clin Epigenetics. [14,17] In contrast, Smith et al. This site needs JavaScript to work properly. Hundred histologically diagnosed cases of HNSCC were studied. Arch Pathol Lab Med. The most important factor in determining the magnitude of gains in sensitivity for CIN2+ (PPA) obtained with p16 IHC appears to be the diagnostic approach an individual ISP takes when interpreting cervical biopsies. and Ai et al., whereas wide variability and discordance was observed when the criteria for grading used were different. Using CPRH&E+p16 as the reference, increase in PPA (sensitivity) and NPA (specificity) within LAST cases was 11.8% (95% CI, 9.5-14.0; P<0.0001) and 9.7% (95% CI, 7.8-11.5; P<0.0001), respectively. no. Results: -, Immunohistochemical expression of epidermal growth factor receptor (EGFR) in South Asian head and neck squamous cell carcinoma: association with various risk factors and clinico-pathologic and prognostic parameters. found no significant association between p16 expression and alcohol consumption. Bethesda, MD 20894, Web Policies [5] Human papillomavirus (HPV) status is associated with p16 expression, and HPV-positive tumors are less likely to harbor p53 mutations. The frequency distributions of the ISPs diagnoses established on H&E-stained slides only (ISPH&E), versus on H&E-stained slides plus p16-stained slides per LAST criteria (ISPH&E+p16/LAST) dichotomized at the CIN2+ (ie, CIN2, CIN3, ACIS, invasive carcinoma)/CIN1 (ie, no CIN, CIN1) threshold are provided in Table 1 for both reference diagnoses, that is CPR on H&E (CPRH&E; Table 1) and on H&E plus p16-stained slides (CPRH&E+p16; Table 1). Smilek P, Dusek L, Vesely K, Rottenberg J, Kostrica R. Correlation of expression of Ki67, EFGR, C-erbB2, MMP-9, p53, bcl2, CD34 and cell cycle analysis with survival in head and neck squamous cell cancer. Accessibility Immunohistochemistry results should be interpreted based on the pattern of positivity. Prevalence of human papillomavirus type 16 DNA in squamous cell carcinoma of the palatine tonsil, and not the oral cavity, in young patients: A distinct clinicopathologic and molecular disease entity. Unauthorized use of these marks is strictly prohibited. Examples of hematoxylin and eosin (H&E) staining and p16INK4a immunohistochemical (p16 IHC) staining, MeSH Results p16 over expression was noted in 22.9% (33 cases), while 21.5% (31 cases) were focal positive and 55.6% (80 cases) were negative for p16 over expression. Risk stratification by. Bookshelf HHS Vulnerability Disclosure, Help Grade is a good prognostic factor associated with distant metastasis of tumors in head-and-neck carcinomas. Correlation of clinicopathological features with immunohistochemical expression of cell cycle regulatory proteins p16 and retinoblastoma: Distinct association with keratinisation and differentiation in oral cavity squamous cell carcinoma. Wald test P-values were calculated comparing the reader-averaged diagnostic performances between ISPH&E and ISPH&E+p16 with the SEs estimated from the bootstrap samples. This increase in the detection of histologic HSIL cases came at the cost of a slight, but statistically significant decrease in NPA (0.8%; 95% CI, 1.1 to 0.5; P<0.0001) when p16 was used in the subset of non-LAST cases in this study (Table 4; Fig. Ventana CINtec Histology uses a mouse monoclonal anti-p16 antibody clone E6H4 to detect mouse p16 INK4A protein in FFPE (CINTtec Histology, Roche, cat. 2008;32:502512. Another study that monitored the impact of adopting the LAST criteria in a large academic laboratory found an approximately one-third reduction in CIN2 diagnoses after their adoption.14 However, other studies have either questioned the usefulness of the LAST criteria, found only a minimal increase in the detection of CIN2,3 using p16 IHC, or reported that the use of p16 IHC resulted in the overdiagnosis of CIN2,3.1517. 11,20,21 Recommendations . See this image and copyright information in PMC. J Low Genit Tract Dis. The use of p16 IHC improves diagnostic agreement in cervical biopsy interpretation 7. Multiplex HPV RNA in situ hybridization/p16 immunohistochemistry: a [22] Ralli et al. J Low Genit Tract Dis. R24 TW007988/TW/FIC NIH HHS/United States, International Agency for Research on Cancer - Screening Group. Stacey M. Gargano MD a , Christopher Sebastiano MD a , Agreement rates of ISPs diagnoses on H&E versus on H&E+p16/LAST with both CPR reference diagnoses dichotomized at the CIN2+ threshold are shown in Table 3. According to Ralli et al., HNSCC was associated more with paan chewing and was more in nonalcoholics. National Library of Medicine The correlation of p16 expression with histologic grading was not found to be statistically significant (P = 0.36). and transmitted securely. PDF Interpretation of p16 Immunohistochemistry In Lower Anogenital Tract Similar to our study, Ralli et al. Am J Clin Pathol. A total of 1100 formalin-fixed, paraffin-embedded cervical punch biopsies representative of a colposcopy referral population in the clinical practice setting was used as the reading set for the study. History of alcohol abuse was present in 24 patients, of which 16 (66.67%) had positive p16 expression. Federal government websites often end in .gov or .mil. Data is temporarily unavailable. The 4 individual reading sets were assigned to the 4 different reader cohorts (1 reading set per reader cohort). 19. 8600 Rockville Pike Routine use of adjunctive. HHS Vulnerability Disclosure, Help 8600 Rockville Pike 2017 May/Jun;25(5):366-373. doi: 10.1097/PAI.0000000000000309. [8] HPV-associated cancers are caused by the expression of HPV's E6 and E7 proteins that bind to and inactivate tumor suppressor proteins p53 and retinoblastoma protein respectively, leading to malignant transformation of HPV infected cells. Thus, it helps to identify patients at high risk for distant metastasis so that the patients at high risk should get efficient systemic treatment. The LAST committee originally estimated that 25% of cervical biopsies would receive p16 IHC if the LAST criteria were followed.12 A lower rate of p16 IHC requests was found by 1 laboratory approximately a year after the LAST criteria were published. The average difference in NPA of ISPs in round 2 (H&E+p16) versus round 1 (H&E only) also differs by diagnostic approach, but moves in the opposite direction to PPA. Kalpien-Meier for epidermal growth factor, Figure 2. Search for Similar Articles Dass SE, Huizenga T, Farshchian M, Mehregan DR. Clin Cosmet Investig Dermatol. BCL-x and bcl2, p53 expression in squamous cell carcinoma of the head and neck cancer. Utility of, 17. Improvements in agreement rates were higher by 60% when CPRH&E+p16 was as reference diagnosis versus CPRH&E: adding p16 per LAST to ISPs slide interpretation resulted in an increase in OPA of 4.4% (95% CI, 3.7-5.0; P<0.0001), PPA of 8.1% (95% CI, 6.5-9.7; P<0.0001), and NPA of 3.5% (95% CI, 2.8-4.2; P<0.0001). Am J Surg Pathol. The whole data were entered in Microsoft Excel Master Sheet and analyzed using Statistical Package for the Social Sciences (SPSS) 15.0 IBM SPSS Statistics, India software. [14] According to studies conducted by Pannone et al., Fregonesi et al., Singhi and Westra and Smith et al., p16 expression was strongly associated with HPV infected HNSCC. Some error has occurred while processing your request. This can be attributed to the habit of tobacco chewing or smoking or alcohol consumption being more common among males in our part of the world which play an important role in the etiopathogenesis of HNSCC. Supported by Ventana Medical Systems Inc (Roche Tissue Diagnostics). p16 antibody IHC staining and interpretation. Please enable scripts and reload this page. For more information, please refer to our Privacy Policy. We conducted a pilot study of whether nonpathologists could accurately diagnose cervical precancer in biopsies using only a basic light microscope, evaluating p16INK4a immunohistochemistry (p16 IHC) of biopsies, and video-based training for both. Some error has occurred while processing your request. An official website of the United States government. FOIA 7. Photographs of these staining patterns have been previously published.11,12,18. Previously we reported an analysis of the CERTAIN study demonstrating that adjunctive p16 IHC significantly improved diagnostic agreement of surgical pathologists by 4.7% when all cervical biopsies are stained.11 The improvement in diagnostic agreement was driven by an 11.5% increase in sensitivity and a 3.0% increase in specificity. The American Journal of Surgical Pathology45(10):1348-1356, October 2021. http://creativecommons.org/licenses/by-nc-nd/4.0/. As HPV integration with the transcription of viral oncoprotein induces overexpression of p16, we can use p16 immunohistochemistry as a surrogate marker of HPV. It was observed that 60 (60%) cases were positive for p16 (inclusive of all grades), while 40 (40%) cases were negative [Figure 5]. When stratified in this manner marked differences were observed in the impact of p16 IHC. Prognostic impact of p16 and p53 expression in oropharyngeal squamous cell carcinomas. 2001;285:15001505. 2017 Mar;39(3):E34-E39. 2011 Jun 7;55(2):e20. Others are quite uncommon types which include lymphoma and adenoid cystic carcinoma. Non-invasive optical biopsy by multiphoton microscopy identifies the live morphology of common melanocytic nevi. Using biopsies collected as part of a screening study conducted in rural China, we randomly selected 50 biopsies with a precancerous diagnosis of cervical intraepithelial neoplasia grade 2 (CIN2) or more severe (CIN2+) and 50 biopsies with diagnosis of CIN less severe than CIN2, and stained them for p16 using a commercial IHC kit. Of note, there was a comparably high increase in PPA when p16 was used within the subset of non-LAST cases (11.0%; 95% CI, 7.8-14.1; P<0.0001) as observed for the subset of the LAST cases (11.8%) (Table 4; Fig. p16 immunohistochemistry in oropharyngeal squamous cell - Nature Nonpathologists can accurately diagnose CIN2+ using p16 immunohistochemistry alone. Compared with the results obtained using hematoxylin and eosin-stained slides only, including p16-stained slides per LAST criteria increased sensitivity and specificity for diagnosing histologic high-grade squamous intraepithelial lesions across all cases by 8.1% (95% confidence interval [95% CI], 6.5-9.7; P<0.0001) and 3.5% (95% CI, 2.8-4.2; P<0.0001), respectively, using expert consensus diagnoses on hematoxylin and eosin+p16 as reference. J Cutan Pathol. The obtained results were analyzed and evaluated using Chi-square test, value of P < 0.05 was taken significant. Head-and-neck squamous cell carcinoma, human papillomavirus, immunohistochemistry, p16. Mills AM, Paquette C, Castle PE, et al. government site. A significant association was seen between p16 and tumor site (P = 0.03) which was in concordance with the study of Yuen et al. Horn LC, Reichert A, Oster A, et al. Epub 2007 May 31. -. is a consultant to Ventana Medical Systems Inc (Roche Tissue Diagnostics). p16INK4a immunohistochemistry in cervical biopsy specimens: A systematic review and meta-analysis of the interobserver agreement. Disclaimer. Interpretation of p16, p53 and mismatch repair protein Disclaimer. cervical biopsy; squamous intraepithelial lesion; immunohistochemistry; p16; LAST; diagnostic agreement. Another third underdiagnosed an average of 10.4% of CIN2+ lesions as normal or CIN1 unless they used p16 IHC and again this was irrespective of whether they thought staining was necessary using the LAST criteria. Data is temporarily unavailable. Singhi AD, Westra WH. Parkin DM, Bray F, Ferlay J, Pisani P. CA Cancer J Clin. P16 - Libre Pathology [12,14], In our study, significant association was seen between p16 expression and sex of the patient (P = 0.004), while in study by Shinohara et al., there was no significant correlation between p16 expression and sex (P = 0.18). Data Sources. Although the results of studies are contradictory, abnormal p16 immunohistochemistry does not reliably predict risk of progression of low grade cervical squamous lesions. . 2021 Aug 24;11(9):1526. doi: 10.3390/diagnostics11091526. Pilloni L, Bianco P, Difelice E, Cabras S, Castellanos ME, Atzori L, Ferreli C, Mulas P, Nemolato S, Faa G. Eur J Histochem. Vaccination programs may provide prevention from HPV infection in high-risk population. [2] More than 90% of HNCs are squamous cell carcinomas (HNSCC). 2015;39:729736. [3] Squamous cell carcinoma of the upper aerodigestive tract typically occurs in older patients in their fifth to seventh decades of life and older. p16 is the most widely studied biomarker in lower anogenital tract squamous intraepithelial lesions and, currently the only recommended biomarker for histological grade assessment. Would you like email updates of new search results? Dragomir LP, Simionescu C, Mrgritescu C, Stepan A, Dragomir IM, Popescu MR. P53, p16 and Ki67 immunoexpression in oral squamous carcinomas. ssues. official website and that any information you provide is encrypted Lpez F, Mkitie A, de Bree R, Franchi A, de Graaf P, Hernndez-Prera JC, Strojan P, Zidar N, Strojan Flear M, Rodrigo JP, Rinaldo A, Centeno BA, Ferlito A. Diagnostics (Basel). doi: 10.7759/cureus.11385. 12. Am J Clin Pathol. 8. Maniar KP, Sanchez B, Paintal A, et al. Using CPRH&E as the reference, improvements were 2.7% (95% CI, 2.0-3.4; P<0.0001) for OPA, 5.1% (95% CI, 3.4-6.9; P<0.0001) for PPA, and 2.3% (95% CI, 1.5-3.1; P<0.0001) for NPA. Oral squamous cell carcinoma is a major cause of death throughout the developed world. One is the cost and logistics of performing p16 IHC on all biopsies. Current Updates on Cancer-Causing Types of Human Papillomaviruses (HPVs) in East, Southeast, and South Asia. From this tissue block, a 4-m-thick slice was cut and its slide was stained according to . The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Second Diagnostic Opinion by Experienced Dermatopathologists in the Setting of a Referral Regional Melanoma Unit Significantly Improves the Clinical Management of Patients With Cutaneous Melanoma. We hypothesized that p16 INK4a immunohistochemistry (p16 IHC) is a sufficiently robust test for CIN2+ that nonpathologists with basic, video-based training on the use of a light microscope and reading of p16 IHC could accurately diagnose CIN2+. Terrell JR, Rybak I, Lyu Y, Konia T, Fung MA, Qi L, Kiuru M. J Cutan Pathol. BMC Res Notes. modify the keyword list to augment your search. Overdiagnosis of HSIL on, 18. Data sources: Scoring of p16INK4a immunohistochemistry based on independent - Nature Readers were grouped into tertiles based on their location on the receiver operating curve, which was estimated by inverse transformation of the linear regression line between the probit of PPA and the probit of 1NPA from all ISPs. No established criteria exist for defining p16 positivity in cytology specimens. p16 is a tumor suppressor protein that acts by slowing the progression from G1 to S phase of the cell cycle. Before El-Mofty SK, Lu DW. Expression of P16INK4a in Uveal Melanoma: New Perspectives. pRb and CyclinD1 Complement p16 as Immunohistochemical Surrogate Markers of HPV Infection in Head and Neck Cancer. Patient with fungating growth on the right lateral border of the tongue. When evaluating the effect of adjunctive p16 within the subset of LAST cases only, both PPA and NPA increased significantly comparing the ISPs reading results without or with p16 to either of the reference diagnoses (Table 4). Adjunctive use of p16 immunohistochemistry for optimizing management of Please enable it to take advantage of the complete set of features! Association of HPV with p16 expression was correlated with a patient's history of abnormal sexual habits like practice of oral sex and with history of multiple sexual partners. Galgano MT, Castle PE, Atkins KA, Brix WK, Nassau SR, Stoler MH. Evaluation of controls, interpretation and reporting of results, and experience of the pathologist are factors accounting for postanalytical variables [44].Because of interobserver variability in reporting p16 IHC, investigators are looking to pursue methods for automated analysis [48].For such an application, the antibody must be easy to use and reproducible with protocols that are . The average difference in NPA in LAST cases was 8-fold greater for the tertile favoring sensitivity over specificity compared with the tertile favoring specificity over sensitivity (16.8% increase in NPA vs. 1.9%, respectively; Table 5). and Ralli et al. Wilson GD, Saunders MI, Dische S, Bentzen SM, Richman PI, Daley FM. Comparison of human papillomavirus in situ hybridization and p16 immunohistochemistry in the detection of human papillomavirus-associated head and neck cancer based on a prospective clinical experience. 2010 Dec;134(12):1785-92. doi: 10.5858/2009-0522-RAR.1. p16 Immunohistochemistry is useful in confirming high-grade squamous intraepithelial lesions (HSIL) in women with negative HPV testing It is believed that almost all squamous cell carcinomas of the cervix are associated with HR-HPV infection. When restricted to LAST cases, the average difference in PPA between when the ISP used p16 versus when they did not was almost 6-fold greater for the tertile favoring specificity over sensitivity compared with the tertile favoring sensitivity over specificity (21.0% increase in PPA vs. 3.6%, respectively; Table 5). One third of the ISPs participating in this study misdiagnosed an average of 21% of CIN2+ lesions as normal or CIN1 unless they used p16 IHC, irrespective of whether they thought staining was necessary using the LAST criteria. 6. A similar distribution of tumors was seen in the study by Boslooper et al.,[18] while the larynx was the most common site of involvement in most of the studies. Patient age and cervical cytology results were available for all cases. They may however show patchy non-block-type staining. Discrepancy in p16 expression in patients with HPV-associated head and Vairaktaris E, Yapijakis C, Psyrri A, Spyridonidou S, Yannopoulos A, Lazaris A, et al. It should be noted that there were fewer diagnoses of CIN2+ made using ISPH&E+p16/LAST compared with ISPH&E, 3773 versus 3996, respectively. Cancer . We would also like to point out that the LAST criteria recommend p16 IHC for all cervical biopsies LSIL, irrespective of their histologic appearance from women with high-risk colposcopic referrals.12 If pathologists decide to stain all cervical biopsies with p16 they should keep in mind many of the other LAST recommendations.12 They should remember that diffuse/block-like p16 positivity is not an independent indicator of HSIL and be careful to not upgrade p16-positive CIN1 lesions to CIN2 or to downgrade histologic CIN3 lesions that are p16 negative. Chen ZW, Weinreb I, Kamel-Reid S, Perez-Ordoez B. Equivocal p16 immunostaining in squamous cell carcinoma of the head and neck: Staining patterns are suggestive of HPV status. In addition, the biopsies underwent 2 rounds of consensus pathology review; once using H&E-stained slides (CPRH&E) and a second time using both H&E-stained and p16-stained slides (CPRH&E+p16). The LAST criteria for p16 IHC were developed based predominantly on expert clinical opinion since at the time they were formulated relatively few studies had analyzed the performance of p16 IHC when applied to specific clinical situations. The site is secure. During the first reading round on H&E-stained slides only, both ISPs as well as the gynecologic pathologists establishing the CPR-derived reference diagnoses were asked whether, based on their review on H&E, they would request adjunctive p16 IHC staining according to LAST criteria as follows: (i) an H&E morphologic differential diagnosis between precancer (CIN2 or CIN3) and a mimic of precancer, (ii) an H&E morphologic interpretation of CIN2 that is or was entertained, and (iii) an H&E morphologic interpretation of CIN1 but at high risk for missed high-grade disease, defined as a prior cytologic interpretation of HSIL, ASC-H, or AGC (not otherwise specified). The readers and the pathologist agreed on p16 IHC scoring for 42 (84%) of the 50 slides of CIN less severe than CIN2 and 37 (74%) of the 50 CIN2+ slides. ROC indicates receiver operator characteristics. The mean sensitivity and specificity of the p16 IHC staining scored by the nonpathologists were 91.7% and 94.1%, respectively, compared to scoring by the pathologist. [12,17] They hypothesized that tumors exhibiting p16 expression effect cell differentiation. HHS Vulnerability Disclosure, Help Journal of Lower Genital Tract Disease19(3):207-211, July 2015. Just as importantly, when p16 IHC was performed according to the LAST criteria, there was an improved correlation between the ISPs diagnosis of CIN2+ and the CPRs diagnosis of CIN2+, using either of the 2 reference diagnoses (CPRH&E or CPRH&E+p16) (Table 1). - A PubMed database search for literature reporting melanocytic lesions and p16 immunohistochemistry was performed. The third tertile has relatively high PPAs and relatively low NPAs compared with the CPR and appears to favor sensitivity over specificity.