Development of alternative photochromic solutions to dual in situ hybridisation - immunohistochemistry assays for evaluating breast and lymphoid neoplasia
National Plan for Research and Development and Innovation for the period 2015-2020 (PNCDI III)
P2 - Increasing the competitiveness of the Romanian economy through RDI: Subprogramme 2.1. Competitiveness through research, development and innovation
Demonstration experimental project (PED) - Competition 2019
Project Director: DR.BOGDAN FETICA
Acronym: DeAlPhISHIC
Code: PN-III-P2-2.1-PED-2019-2308
Promoter: INSTITUTE OF ONCOLOGY "PROF.DR.I.CHIRICUTA" CLUJ-NAPOCA
The Contracting Authority: Executive Unit for Financing Higher Education, Research, Development and Innovation - UEFISCDI
Contract no.: 388PED / 2020
Total Buget: 600.000 RON
Public Buget: 600.000 RON
Duration: 01.11.2020 - 31.10.2022
Prezentarea sintetică a rezultatelor obținute în cadrul proiectului
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Project Description
The scope of this project is to develop an alternative method to dual immunohistochemistry – in situ hybridization assays applied in breast and hematopoietic neoplasia, by incorporating the use of photochromic dyes, that can create a negative imprint image of the IHC assays on slides, which also represents the demonstration model of our project. We considered the following objectives for achieving the scope of the project:
Objective 1. Investigation of the FICTION technique (Fluorescence Immuno-phenotyping and Interphase Cytogenetics as a Tool of the Investigation of Neoplasm) for the assessment of the HER2 status in breast invasive carcinomas by comparing it to the current gold standard (sequential HER2 testing by immunohistochemistry (IHC) and in situ hybridization(ISH)).
Objective 2. Development of an alternative protocol to the dual IHC-ISH assays that incorporates the use of photochromic dyes for the assessment of breast and lymphoid neoplasia.
Objective 3. Validation of the photochromic dyes protocol for assessing lymphoid and breast neoplasia, by comparing it to the current gold standard (sequential testing by IHC and ISH in HER2), to double automated IHC stains and to the validated FICTION technique.
Publications related to the project
Gitr-Expressing Regulatory T-Cell Subsets are Increased in Tumor-Positive Lymph Nodes from Advanced Breast Cancer Patients as Compared to Tumor-Negative Lymph Nodes.
Krausz LT, Fischer-Fodor E, Major ZZ, Fetica B.
International Journal of Immunopathology and Pharmacology, 25(1):59-66, 2012. doi: 10.1177/039463201202500108
Differential Peripheral Blood Gene Expression Profile Based on Her2 Expression on Primary Tumors of Breast Cancer Patients.
Tudoran O, Virtic O, Balacescu L, Pop L, Dragla F, Eniu A, Fetica B et al.
PLoS ONE, 9(7): e102764, 2014. doi: 10.1371/journal.pone.0102764
Blood Genome-Wide Transcriptional Profiles of HER2 Negative Breast Cancers Patients.
Balacescu O, Balacescu L, Virtic O, Visan S, Gherman C, Drigla F, Pop L, Bolba-Morar G, Lisencu C, Fetica B, Tudoran O, Berindan-Neagoe I.
Mediators of Inflammation, vol. 2016, Article ID 3239167, 12 pages, 2016. doi: 10.1155/2016/3239167
Classification of non-Hodgkin lymphoma in South-eastern Europe: review of 632 cases from the international non-Hodgkin lymphoma classification project.
S Dotlic, AM Perry, G Petrusevska, B Fetica, J Diebold, KA MacLennan, HK Müller-Hermelink, BN Nathwani, E Boilesen, M Bast, JO Armitage, DD Weisenburger
British Journal of Haematology, 171, 366–372, 2015. doi: 10.1111/bjh.13586
Non-Hodgkin lymphoma in Romania: a single-centre experience.
B Fetica, P Achimas-Cadariu, B Pop, D Dima, L Petrov, AM Perry, BN Nathwani, HK Müller-Hermelink, J Diebold, KA MacLennan, A Fulop, ML Blaga, D Coza, A Irimie, DD Weisenburger
Hematological Oncology, 35: 198–205, 2017. doi: 10.1002/hon.2266
High prevalence of viral hepatitis in a series of splenic marginal zone lymphomas from Romania.
B Fetica, B Pop, ML Blaga, A Fulop, D Dima, MT Zdrenghea, CI Vlad, AS Bojan, P Achimas-Cadariu, CI Lisencu, A Irimie, DD Weisenburger
Blood Cancer Journal , volume 6, page e498, 2016. doiI: 10.1038/bcj.2016.102
Context
Breast cancer is the most common type of cancer diagnosed in women and is also the main cancer related death cause.
Hormone receptors expression and the HER2 status are the only validated predictive factors and their determination is used to guide endocrine
and trastuzumab targeted therapy. Currently two methods are employed for the determination of the HER2 status: immunohistochemistry (IHC),
used to evaluate protein expression and in situ hybridization (ISH), used to determine the HER2 gene copies number. Fluorescence in situ
hybridization (FISH) is considered the gold standard. The high incidence, prevalence and mortality rates have driven the need for research in this field.
At the other end of the incidence spectrum lymphoid neoplasms represent a distinct category of neoplasms, which pose a great deal of challenges
for clinicians, pathologist, researchers and other professional categories that share an interest in the field. The study of
lymphoid neoplasia represents a research field in which there is a need for improving on diagnostic techniques and on developing
means for identifying molecular changes in tumor cells, with application in targeted therapy development and administration.
The latest Classification of Hematopoietic Neoplasms and Lymphoid Tissue evolved into a real global consensus on hematologic
neoplasms and introduced ancillary techniques in the classification of lymphomas in the context of morphology and clinical data
(immunophenotypic, molecular biology, cytogenetic).
Objectives
The project aims to develop an alternative method to dual
IHC – ISH assays applied in breast and hematopoietic neoplasia, by incorporating the use of photochromic dyes, that can create a negative imprint
image of the IHC assays on slides, which also represents the demonstration model of our project.
Specific objective 1: Investigation of the FICTION technique (Fluorescence Immuno-phenotyping and Interphase Cytogenetics as a Tool of the
Investigation of Neoplasm) for the assessment of the HER2 status in breast invasive carcinomas, in the current context of the 2018 HER2 Testing in
Breast Cancer, by comparing it to the current gold standard (sequential HER2 testing by immunohistochemistry (IHC) and in situ hybridization(ISH)).
Specific objective 2: Development of an alternative protocol to the dual IHC-ISH assays that incorporates the use of photochromic dyes for
the assessment of breast and lymphoid neoplasia.
Specific objective 3: Validation of the photochromic dyes protocol for assessing lymphoid and breast neoplasia, by comparing it to the
current gold standard (sequential testing by IHC and ISH in HER2), to double automated IHC stains and to the validated FICTION technique.
Impact
Achieving objective 1 translates into providing an alternative method to sequential HER2 testing by IHC and ISH, that allows simultaneous
assessment of protein expression and gene status with superior capabilities in identifying areas of interest on slides, that preserve
the tissue in biopsy specimens, by reducing the need for supplementary tissue sections. Photochromic dyes have gained attention in the
field of life sciences due to an ongrowing demand for controllable molecular switches and in other industry fields like optical circuitry
(switches and logic gates), information technology and nanotechnology. The development of a photochromic dye protocol (objective 2) will
result in a method that creates a negative imprint of the IHC staining pattern on the glass slides coated with photochromic dyes.
To our best knowledge the application of photochromic dyes for improving brightfield microscopy has never been attempted.
By developing the photochromic dyes protocol, we want to provide a laboratory validated method (objective 3) that can be used in various
combinations with IHC and ISH, for concomitant in situ assessments.
The project uses basic IHC and ISH techniques, which in different combinations represent a TRL 3 starting point,
according to the description provided in the information package, and evolves to TRL 4 by validating in a laboratory environment
the proposed concept, a method that incorporates photochromic dyes into current IHC/ISH protocols and functions as a surrogate for
the IHC technique in dual assays.
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