
==== Front
F1000Res
F1000Res
F1000Research
2046-1402
F1000 Research Limited London, UK

10.12688/f1000research.134757.2
Study Protocol
Articles
Protocol for correlation of histological risk assessment/scoring system with a depth of invasion in oral squamous cell carcinoma
[version 2; peer review: 2 approved]

Sonone Archana Conceptualization Formal Analysis Funding Acquisition Investigation Methodology Project Administration Resources Software Supervision Validation Visualization Writing – Original Draft Preparation Writing – Review & Editing https://orcid.org/0000-0001-9225-7919
a1
Hande Alka Supervision Validation Visualization https://orcid.org/0000-0002-8576-7897
1
Pakhale Aayushi Supervision Validation Visualization https://orcid.org/0000-0002-0529-3856
1
Gawande Madhuri Supervision Validation Visualization 1
Patil Swati Supervision Validation Visualization 1
1 Department of Oral & Maxillofacial Pathology and MicrobiologySharad Pawar Dental College & Hospital, Datta Meghe Institute of Higher Education, Wardha, Maharashtra, 442004, India
a archana.oralpath@dmiher.edu.in
No competing interests were disclosed.

31 5 2024
2023
12 132629 5 2024
Copyright: © 2024 Sonone A et al.
2024
https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Introduction

The commonest type of cancer in the head and neck region is oral squamous cell carcinoma (OSCC) due to its high rates of occurrence and mortality. The early diagnosis of oral cancer gives better prognosis. Brandwein-Gensler criteria predict the early stage of OSCC cases with a high risk of locoregional recurrence.

Objectives

To correlate Brandwein-Gensler criteria and depth of invasion of OSCC with three-year survival.

Methodology

In the study, This study will include 80 random histopathologically-diagnosed cases of OSCC. hematoxylin-eosin (HE)-stained section slides will be used to evaluate, Brandwein and Gensler criteria by three histopathologists in a blinded manner. The depth of invasion assessment will be done from the basement-membrane (BM), in regions where the BM has been lost, as well as from an illustrative line connecting the BM from the neighbouring epithelium to the point of deepest tumour invasion in the connective-tissue stroma with the help of a research microscope (Leica-DMLB2) in resected tissue specimens of OSCC cases.

Expected results

The present study will find the correlation between Brandwein-Gensler criteria and depth of invasion in OSCC in order to evaluate the locoregional recurrence in OSCC cases. In high-risk OSCC cases, there may be an increased depth of invasion in resected tissues.

Conclusions

We hypothesized that the correlation between Brandwein-Gensler criteria and depth of invasion can be used as an independent predictor for locoregional recurrence in OSCC.

Oral squamous cell carcinoma
Brandwein Gensler Criteria
Depth of Invasion
The author(s) declared that no grants were involved in supporting this work.Revised Amendments from Version 1

In the new version, we have updated the methodology in detail.
==== Body
pmcIntroduction

Oral squamous cell carcinoma (OSCC) is the most widely occurring cancer in the head-neck region. 1 In 2018, OSCC claimed around 177,000 global fatalities. 2 Despite of the advanced treatment modalities and therapeutic approaches, the overall survival rate in OSCC did not rise by more than 50% during a five-year period. 3 The clinical diagnosis of OSCC is confirmed by histopathology. In the adjunct therapy such as chemotherapy radiotherapy, histopathological report of OSCC is important. Due to this, many studies emphasize on the histopathological parameters of resected oral cancer tissues in the pathology reports. 4 Several authors have proposed various grading systems for OSCC. The first grading system developed by Broder (1920), which is still advised by the WHO 5 , 6 is likely the most well-known prognostication method using a subjective evaluation of significant histopathological features like differentiation degree of tumor cells, cellular pleomorphism, and mitotic activity, and is grouped into WDSCC (well differentiated), MDSCC (moderately differentiated) and PDSCC (poorly differentiated). Although MDSCC comprises up to 90% of oral cancers, this approach, while widely identified and used, still has poor discriminatory value because in MDSCC pathologically there is a poor cellular differentiation than the WDSCC. 6 , 7 Annoreth et al. gave another classification which emphasise on the association between the tumour and adjacent tissue. 8 This included aspects like the leukocyte invasion and the degree of pattern and stage of invasion. A method of invasive-tumour-front grading (IFG) comprising five histopathological characteristics involving lymphocyte-host-response (LHR) was later devised by Bryne et al. 9 Because of the small sample sizes, varied locations of the tumour, and examination of various types of specimens, these models have not proved successful. A risk assessment model put forth in 2005 by Brandwein-Gensler and colleagues was said to have better prognostic value than previously mentioned systems. 10 It includes the combined assessment of three important histopathological parameters: PNI (perineural-invasion), LHR (lymphocyte-host-response), and WPOI (worst-pattern of invasion). Scoring is done for all three parameters individually and then the sum of the three variables is computed. The Brandwein-Gensler (BG) system showed a significant correlation with the patient’s locoregional recurrence and overall survival, especially in low-stage oral cancers. More aggressive characteristics receive weighted point values in the model. 11 Adjuvant radiotherapy may be beneficial in cases of high-risk, low-stage oral cancers, even when the margins are satisfactory. The TNM classification for OSCC (eight edition, 2017), emphasizes on depth-of-tumour invasion (DOI) along with largest diameter of the tumour to determine the T-stage 12 and signifies a shift in the paradigm in Oral Pathology. When determining the specific course of treatment for a particular case, a combined evaluation of clinical staging and histological grading may be a more reliable method. 1 The main goal of our study is to determine the association of DOI with the histological risk assessment/scoring system in order to assess the combined effects of these factors on OSCC pathological staging and locoregional recurrence in three-year survival rate.

Methods

Trial design

This study will be retrospective, using archival tissue from OSCC cases.

Study setting

This cross-sectional study will be carried out at Sharad Pawar Dental College and Hospital (SPDC&H), Datta Meghe Institute of Higher Education, Sawangi-Meghe, Wardha in the Oral Pathology and Microbiology Department. This study will include 80 random histopathologically-diagnosed cases of OSCC. Computer-generated randomization technique was used. The staging will be carried out on the basis of the American Joint Committee on Cancer (AJCC) eighth edition criteria. 13 Cases with a previous head-neck cancer history, pre-surgical radiation therapy, chemotherapy or any surgery (apart from biopsy) and recurrent or distant disease will be excluded. Patient data comprising of age and sex of the patient, site of the lesion, clinical appearance of the tumor, habits including tobacco and alcohol use and status of lymph node will be obtained.

Intervention

Staining of the archival slides and new slides (where needed) will be done with Heamtoxyline and eosin stain. Worst-pattern of invasion WPOI, Lymphocyte-host-response LHR, and Perineural-invasion PNI will be assessed by three histopathologists in a blinded manner. Evaluation will be done according to the Risk Model mentioned below. Three groups will be made depending on the sum of the score of each case:

Group 1 – low-risk cases = total score 0

Group 2 – moderate-risk cases = 1 or 2

Group 3 – high risk-cases = 3 or more.

The DOI assessment will be done from the basement-membrane (BM), in regions where the BM has been lost, as well as from an illustrative line connecting the BM from the neighbouring epithelium to the point of deepest tumour invasion in the connective-tissue stroma with the help of a research microscope (Leica-DMLB2) with standard software (Leica-Q-win). The DOI will be measured and recorded.

Risk assessment system derived from Brandwein-Gensler et al.

WPOI:

WPOI-1: Broad-pushing borders – Score 0

WPOI-2: Broad-pushing fingers – Score 0

WPOI-3: Large tumour cells islands (with more than fifteen tumour cells in each island) – Score 0

WPOI-4: Small islands of tumour (less than fifteen tumour cells in each island) – Score +1

WPOI-5: Satellites of tumour cells, at ≥1mm distance from the major tumour – Score +3

Lymphocytic-host-response (LHR):

LHR-1 (strong): Dense rim of lymphoid infiltrate surrounding the tumour at the advancing edge/invasive front – Score 0

LHR-2 (intermediate): patches of lymphoid infiltrate in few regions – Score +1

LHR-3 (weak): Presence of very little or no lymphoid infiltrate, no patches seen – Score +3

Perineural-invasion:

No invasion – Score 0

Small nerves: Tumour cells surrounding small nerves – Score +1

Large nerves: Tumour cells surrounding nerves, ≥1 mm in diameter – Score +3

Outcomes

The present study will find the correlation between Brandwein-Gensler criteria and DOI in OSCC. In the high-risk category, there may be increased DOI in resected tissue of OSCC.

Sample size

Considering the prevalence of OSCC as 70.7% in the outpatient department of the Oral Pathology and Microbiology department, using the single proportion formula, sample size is calculated by applying the formula: n≥Z21−α/2∗p∗1−p/d2

Where,

Z21−α/2 - The significance level at 5%

i.e. 95 % confidence interval = 1.96

p = Sample showing positive CD44 expression focally in small group cells in the basal layer of epithelium = 70.7% = 0.707

E=Error of margin=10%=0.10

n=1.962×0.707×1−0.707/0.102

n = 80

Dissemination

The results will be published in an indexed journal.

Study status

The study has not started yet.

Discussion

Incidence of OSCC in Indian men (11.28%) ranks first while it ranks fifth in women (4.3%). 1 , 2 The clinical diagnosis is always confirmed by the histopathology. There are numerous histopathological grading systems mentioned in the literature for OSCC. Among them, the BG risk predictive model gives the best results regarding the loco-regional recurrence of OSCC. 4

Arun Chaturvedi evaluated the BG risk predictive model in surgically treated OSCC patients in North India. The author studied 149 patients with histologically diagnosed OSCC. The patients were divided into three groups: low, moderate, and high risk according to the BG criteria. Out of 149 patients, 17 patients showed locoregional recurrences (11.4%). Most of these 17 patients belonged to the high-risk category of the BG risk model. The authors hypothesised that the BG risk model is predictive of locoregional recurrences for OSCC undergoing primary surgery and it can be used to model for recognition of recurrences and prevention at early stages of the disease. 14

Rhayany de Castro Ribeiro Lindenblatt studied 53 cases of OSCC in which they evaluated four grading systems (Malignancy Grading of the Deep Invasive Margins, Multiparameter Grading System, Histologic Risk Assessment and the World Health Organization grading system). They concluded that the Histologic Risk Assessment scoring system established the best results for locoregional recurrence and survival prediction in OSCC patients. 15

Naomi Rahman evaluated the BG risk model criteria in OSCC patients to assess disease progression and survival of patients. They studied 134, OSCC cases, in which they compared two AJCC criteria, i.e. seventh and eighth. In the latest eighth criteria they included DOI according to which they staged the tumor. Due to this, 30 cases of OSCC previously classified into the T1 and T2 stage according to the seventh edition were upstaged to T3 stage according to the eighth AJCC criteria. Three individual histopathologists scored BG risk model criteria and DOI of the same samples. They concluded that the BG is the most effective parameter for grading tumors and locoregional recurrence of patients. 16

The staging of OSCC, which is based on the TNM system, has been used for several years to estimate both the clinical response to therapy and survival outcomes. In the eighth AJCC edition, DOI is included as a new histopathological parameter for the assessment of tumor grade, The DOI is measured from the basement membrane to invasive the front of the tumor. The stage and grade of the tumor are major contributing factors to prognosis and treatment selection for OSCC. 17 As the BG risk model gives the locoregional recurrence status of OSCC, assessment and corelation of BG model and depth of tumor give the best results in predicting three-year survival in locoregional recurrence of OSCC patients.

Conclusions

For OSCC, the BG risk model comprises effective criteria to determine locoregional recurrence. Therefore, it can be used in early-stage OSCC for identification and prevention of the recurrences. The correlation between BG and DOI in resected OSCC tissues will give better results about the locoregional recurrence and survival rate of OSCC.

Ethical considerations

Ethical approval was received from Datta Meghe Institute of Higher Education and Research, Sawangi, Wardha, Maharashtra India (IEC reference number- DMIHER (DU)/IEC/2023/578).

Acknowledgements

The authors would like to thank their institute and colleagues.

Data availability

Underlying data

No data are associated with this article.

Reporting guidelines

Zenodo: STROBE checklist for “Protocol for correlation of histological risk assessment/scoring system with a depth of invasion in oral squamous cell carcinoma”, https://doi.org/10.5281/zenodo.7895574 . 18

Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).

10.5256/f1000research.167199.r314720
Reviewer response for version 2
Hardillo Jose A. 1Referee
1 Erasmus MC Cancer Institute, Rotterdam, The Netherlands
9 9 2024 Copyright: © 2024 Hardillo JA
2024
https://creativecommons.org/licenses/by/4.0/ This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Version 2recommendationapprove
I have read the revised form of this protocol for correlation of histological risk assessment/scoring system with a depth of invasion in oral squamous cells carcinoma well. I believe that this will be a good and relevant study. I do not have major concerns regarding the protocol except that the authors should clearly define that they are going to include only T1N0 and T2N0 tumors.

Is the study design appropriate for the research question?

Yes

Is the rationale for, and objectives of, the study clearly described?

Yes

Are sufficient details of the methods provided to allow replication by others?

Yes

Are the datasets clearly presented in a useable and accessible format?

Yes

Reviewer Expertise:

head and neck tumor surgery, oligometastasis

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

10.5256/f1000research.167199.r285383
Reviewer response for version 2
Polizzi Alessandro 1Referee https://orcid.org/0000-0001-6717-8899

1 University of Catania, Catania, Italy
8 7 2024 Copyright: © 2024 Polizzi A
2024
https://creativecommons.org/licenses/by/4.0/ This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Version 2recommendationapprove
In their revised version the authors successfully addressed my comments. The protocol may be accepted after the following minor correction:

_ ABSTRACT, Correct the errors in the sentences: “In the study, This study will include 80 random histopathologically-diagnosed cases of OSCC. hematoxylin-eosin…”.

Is the study design appropriate for the research question?

Yes

Is the rationale for, and objectives of, the study clearly described?

Yes

Are sufficient details of the methods provided to allow replication by others?

Partly

Are the datasets clearly presented in a useable and accessible format?

Not applicable

Reviewer Expertise:

Oral pathology and medicine; periodontology, orthodontics, oral surgery.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

10.5256/f1000research.147837.r261195
Reviewer response for version 1
Polizzi Alessandro 1Referee https://orcid.org/0000-0001-6717-8899

1 University of Catania, Catania, Italy
12 4 2024 Copyright: © 2024 Polizzi A
2024
https://creativecommons.org/licenses/by/4.0/ This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Version 1recommendationapprove-with-reservations
In the manuscript entitled “Protocol for correlation of histological risk assessment/scoring system with a depth of invasion in oral squamous cell carcinoma” the authors aimed to present a protocol to correlate Brandwein-Gensler criteria and depth of invasion of OSCC with three-year survival.

Here are reported some suggestions to improve the quality of the manuscript:

ABSTRACT: Please, give more details about the methodology and study design in the abstract.

INTRODUCTION: The authors reported that “Although MDSCC comprises up to 90% of oral cancers, this approach, while widely identified and used, still has poor discriminatory value”. Please, better explain why it has poor discriminatory value.

“The Brandwein-Gensler (BG) system showed a significant correlation with the patient’s locoregional recurrence and overall survival, especially in low-stage oral cancers”. Please, give more details about the results you found in the literature about the strengths of this correlation related to recurrence and overall survival.

 “The TNM classification for OSCC (eigtth edition…”. Please correct the error.

METHODS:  Please better explain if this study will be a retrospective or a cross-sectional study.

The authors cited the AJCC eighth edition criteria. Please explain the acronym and add a reference.

Please, give more details about the randomized selection of histopathologically-diagnosed cases of OSCC.

“Staining of the archival slides and new slides (where needed) will be done with HE. WPOI, LHR, and PNI will be assessed by three histopathologists in a blinded manner”. Please specify all the acronyms the first time they have been cited.

Is the study design appropriate for the research question?

Yes

Is the rationale for, and objectives of, the study clearly described?

Yes

Are sufficient details of the methods provided to allow replication by others?

Partly

Are the datasets clearly presented in a useable and accessible format?

Not applicable

Reviewer Expertise:

Oral pathology and medicine; periodontology, orthodontics, oral surgery.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

Sonone Dr. Archana Oral Pathology & Microbiology, Sharad Pawar Dental Collge and Hospital sawangi meghe wardha, Wardha, Maharashtra, India

28 5 2024 Dear Reviewer, 

Thank you for your response , I have done all the corrections which were mentioned in the comment. 

The Brandwein-Gensler (BG) system showed a significant correlation with the patient’s locoregional recurrence and overall survival, especially in low-stage oral cancers”. Please, give more details about the results you found in the literature about the strengths of this correlation related to recurrence and overall survival. meaning of this sentence- this point elaborated in the discussion section .

Thank you regards,

Dr. Archana Sonone

Competing interests: No competing interests were disclosed.

Competing interests: No competing interests were disclosed.

Competing interests: No competing interests were disclosed.

Competing interests: NO
==== Refs
References

1 Dikshit R : Cancer mortality in India: a nationally representative survey. Lancet. 2012;379 (9828 ):1807–1816. 10.1016/S0140-6736(12)60358-4 22460346
2 Stewart BW Wild CP : World Cancer Report 2014. Lyon: International Agency for Research on Cancer;2014.
3 Li Y Bai S Carroll W : Validation of risk model: high risk classification and tumor pattern of invasion predict outcome for patients with low-stage oral cavity squamous cell carcinoma. Head Neck Pathol. 2013;7 :211–223. 10.1007/s12105-012-0412-1 23250819
4 Brandwein-Gensler M Teixeira MS Lewis CM : Oral squamous cell carcinoma: histologic risk assessment, but not margin status, is strongly predictive of local disease free and overall survival. Am J Surg Pathol. 2005;29 (2 ):167–178. 10.1097/01.pas.0000149687.90710.21 15644773
5 Massano J Regateiro FS Januario G : Oral squamous cell carcinoma: review of prognostic and predictive factors. Oral Surg Oral Med Oral Pathol Oral RadiolEndod. 2006;102 (1 ):67–76. 10.1016/j.tripleo.2005.07.038
6 Zevallos JP Mazul AL Walter V : Gene expression subtype predicts nodal metastasis and survival in human papillomavirus-negative head and neck cancer. Laryngoscope. 2019;129 :154–161. 10.1002/lary.27340 30247749
7 Bobdey S Sathwara J Jain A : Squamous cell carcinoma of buccal mucosa: An analysis of prognostic factors. South Asian J Cancer. 2018 Jan-Mar;7 (1 ):49–54. 10.4103/sajc.sajc_317_16 29600236
8 Brandwein-Gensler M Smith RV Wang B : Validation of the histological risk model in a new patient cohort with primaryhead and neck squamous cell carcinoma. Am J Surg Pathol. 2010;34 :676–688. 10.1097/PAS.0b013e3181d95c37 20414102
9 Vered M Dayan D Dobriyan A : Oral tongue squamous cell carcinoma: recurrent disease is associated with histopathologic risk score and young age. J Cancer Res Clin Oncol. 2010;136 :1039–1048. 10.1007/s00432-009-0749-3 20054559
10 Jakobsson PA Eneeroth CM Killander D : Histologic classifcation and grading of malignancy in carcinoma of the larynx. Acta Radiol Ther Physiol. 1973;12 :1–8.
11 Lo WL Ko S-Y Chi LY : Outcomes of oral cancer in Taiwan after surgical therapy: factors afecting survival. J Oral Maxillofac Surg. 2003;61 :751–758. 10.1016/S0278-2391(03)00149-6 12856245
12 Gonzales-Moles MA Esteban F Rodriguez-Archilla A : Importance of tumourthickness measurement in prognosis of tongue cancer. Oral Oncol. 2002;38 :394–397. 10.1016/S1368-8375(01)00081-1 12076706
13 Amin MB Greene FL Edge SB : The Eighth Edition AJCC Cancer Staging Manual: Continuing to build a bridge from a population-based to a more “personalized” approach to cancer staging. CA Cancer J Clin. 2017 Mar;67 (2 ):93–99. 10.3322/caac.21388 28094848
14 Chaturvedi A Husain N Misra S : Validation of the Brandwein Gensler Risk Model in Patients of Oral Cavity Squamous Cell Carcinoma in North India. Head Neck Pathol. 2020 Sep;14 (3 ):616–622. 10.1007/s12105-019-01082-6 31552620
15 Lindenblatt RC Martinez GL Silva LE : Oral squamous cell carcinoma grading systems--analysis of the best survival predictor. J Oral Pathol Med. 2012 Jan;41 (1 ):34–39. 10.1111/j.1600-0714.2011.01068.x 21902722
16 Rahman N MacNeill M Wallace W : Reframing Histological Risk Assessment of Oral Squamous Cell Carcinoma in the Era of UICC 8th Edition TNM Staging. Head Neck Pathol. 2021 Mar;15 (1 ):202–211. Epub 2020 Jul 13. 10.1007/s12105-020-01201-8 32661668
17 Shah JP Montero PH : New AJCC/UICC staging system for head and neck, and thyroid cancer. Revista Médica Clínica Las Condes. 2018;29 (4 ):397–404. 10.1016/j.rmclc.2018.07.002
18 Sonone A Dr. Hande A Dr. Pakhale A Dr. : Protocol For Correlation Of Histological Risk Assessment/Scoring System With Depth Of Invasion In Oral Squamous Cell Carcinoma. Zenodo. 2023. 10.5281/zenodo.7895574
