Modeling Oral-Esophageal Squamous Cell Carcinoma in 3D Organoids

Samuel Flashner; Cecilia Martin; Norihiro Matsuura; Masataka Shimonosono; Yasuto Tomita; Masaki Morimoto; Ogoegbunam Okolo; Victoria X. Yu; Anuraag S. Parikh; Andres J. P. Klein-Szanto; Kelley Yan; Joel T. Gabre; Chao Lu; Fatemeh Momen-Heravi; Anil K. Rustgi; Hiroshi Nakagawa

doi:10.3791/64676

Modelowanie raka płaskonabłonkowego jamy ustnej i przełyku w organoidach 3D

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

Modeling Oral-Esophageal Squamous Cell Carcinoma in 3D Organoids

Modelowanie raka płaskonabłonkowego jamy ustnej i przełyku w organoidach 3D

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10:43 min

December 23, 2022

DOI: 10.3791/64676-v

Samuel Flashner*¹, Cecilia Martin*^1,2, Norihiro Matsuura¹, Masataka Shimonosono¹, Yasuto Tomita¹, Masaki Morimoto¹, Ogoegbunam Okolo¹, Victoria X. Yu^1,3, Anuraag S. Parikh^1,3, Andres J. P. Klein-Szanto⁴, Kelley Yan^1,2, Joel T. Gabre^1,5, Chao Lu^1,6, Fatemeh Momen-Heravi^1,7, Anil K. Rustgi^1,5, Hiroshi Nakagawa^1,2,5

¹Herbert Irving Comprehensive Cancer Center,Columbia University, ²Organoid and Cell Culture Core, Columbia University Digestive and Liver Diseases Research Center,Columbia University, ³Department of Otolaryngology, Head and Neck Surgery,Columbia University, ⁴Histopathology Facility,Fox Chase Cancer Center, ⁵Division of Digestive and Liver Diseases, Department of Medicine,Columbia University, ⁶Department of Genetics and Development,Columbia University, ⁷Section of Oral, Diagnostic and Rehabilitation Sciences, College of Dental Medicine,Columbia University

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Overview

This protocol describes the generation and characterization of murine oral-esophageal 3D organoids that model normal, preneoplastic, and squamous cell carcinoma lesions. These organoids serve as a platform for studying esophageal squamous cell carcinoma (ESCC) and its molecular changes.

Key Study Components

Area of Science

Neuroscience
Oncology
Cell Biology

Background

Esophageal squamous cell carcinoma (ESCC) is a prevalent and deadly cancer worldwide.
3D organoids can accelerate research in ESCC.
Single cell-derived organoids capture the genetic diversity of tumors.
These models are useful for testing new therapeutic strategies.

Purpose of Study

To create a model that mimics ESCC for research purposes.
To understand the molecular changes during ESCC initiation and development.
To provide a platform for functional annotation of these changes.

Methods Used

Utilization of 4NQO to induce ESCC in mice.
Isolation of single cell-derived 3D organoids.
Characterization of organoids for functional studies.
Preparation of organoids for paraffin embedding.

Main Results

Organoids represent the genetic heterogeneity of ESCC.
They provide a relevant physiological model for testing therapies.
Demonstrated techniques for organ collection and preparation.
Facilitated understanding of tumorigenesis in ESCC.

Conclusions

3D organoids are valuable tools for ESCC research.
They help elucidate the molecular mechanisms of cancer development.
This model can aid in the identification of therapeutic targets.

Frequently Asked Questions

What is the significance of using 3D organoids?

3D organoids provide a more physiologically relevant model for studying cancer compared to traditional 2D cultures.

How are the organoids generated?

Organoids are generated from single cells derived from mice treated with 4NQO, which induces ESCC.

What are the applications of these organoids?

They can be used to test novel therapeutic strategies and study the genetic changes in ESCC.

Who contributed to the study?

Yasuto Tomita and Norihiro Matsuura contributed to the demonstration of organ collection and preparation techniques.

What is the main focus of this research?

The main focus is to understand the molecular changes associated with ESCC using a 3D organoid model.

Why is ESCC a significant health concern?

ESCC is prevalent and has a high mortality rate, making it a critical area of research in oncology.

Ten protokół opisuje kluczowe kroki do wygenerowania i scharakteryzowania mysich organoidów 3D ustno-przełykowych, które reprezentują normalne, przednowotworowe i płaskonabłonkowe zmiany rakowokomórkowe wywołane przez chemiczną kancerogenezę.

Rak płaskonabłonkowy przełyku, ESCC, jest śmiertelny i powszechny na całym świecie. Trójwymiarowe organoidy mogą być wykorzystane do przyspieszenia postępu w tej dziedzinie w celu zwalczania poważnego obciążenia ESCC. Organoidy 3D pochodzące z pojedynczych komórek od myszy leczonych 4NQO mogą być łatwo i niedrogo manipulowane w celu funkcjonalnej adnotacji zmian molekularnych towarzyszących inicjacji i rozwojowi ESCC.

Model ten pozwala uchwycić niejednorodność genetyczną nowotworów wywołanych mutagenem. W związku z tym organoidy te stanowią fizjologicznie istotną platformę do testowania nowych strategii terapeutycznych lub identyfikowania istotnych zmian genetycznych napędzających nowotworzenie nowotworów. W demonstracji sekcji zwierząt i pobierania narządów pomoże Yasuto Tomita, współpracownik personelu, a przygotowanie organoidu do procedury zatapiania parafiny zademonstruje Norihiro Matsuura, doktor habilitowany z mojego laboratorium.

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