Construction of micropapillary lung adenocarcinoma organoids and screening of targeted drugs

doi:10.11958/20231326

Abstract

Abstract:

Objective To establish a culture method for micropapillary lung adenocarcinoma organoids and conduct targeted drug screening. Methods Organoids were extracted and cultured from a surgical tissue sample of a patient diagnosed with micropapillary lung adenocarcinoma, and the growth of lung cancer organoids was observed and recorded dynamically. The morphological and gene expression characteristics of tumor cells between lung cancer organoids and parental tissue were compared using hematoxylin eosin (HE) staining and immunohistochemical methods. Real time fluorescence quantitative polynucleotide chain reaction (qRT-PCR) method was used to detect gene mutations in lung cancer parental tissue and organoids. Finally, based on results of genetic testing, targeted drugs were selected and their therapeutic effects were verified. Results We have successfully cultured spherical organoids from micropapillary lung adenocarcinoma tissue, which can be passaged for at least 3 generations. HE staining results showed that the morphology of tumor cells in organoids was roughly consistent with that of parental tissue. The immunohistochemical results showed that the protein expression levels of various genes in lung cancer organoids and parental tissue were roughly the same. Results of gene mutation analysis showed that the mutated genes in lung cancer parental tissue and organoids were consistent, both reflecting RET fusion. The screening results of targeted drugs based on lung cancer organoids showed that vandertinib had the best anti-tumor effect in vitro. Conclusion Drug screening experiments based on micropapillary lung adenocarcinoma organoids can screen highly efficient targeted drugs in a short period of time, which may benefit patients with micropapillary lung adenocarcinoma.

Key words: adenocarcinoma of lung, organoids, drug screening assays, antitumor, targeting preparation, vandetani

CLC Number:

R734.2

Figures/Tables 5

Fig.1 Culture of micropapillary lung adenocarcinoid organoid

Fig.2 HE staining and immunohistochemistry experiments identify the phenotype of micropapillary lung adenocarcinoma organoid

Tab.1 Analysis of immunohistochemical result score

抗体	肿瘤亲本组织			类器官
抗体	阳性细胞占比评分	染色强度评分	总评分	阳性细胞占比评分	染色强度评分	总评分
TTF-1	3	3	9（高表达）	3	2	6（高表达）
CK7	4	3	12（高表达）	4	3	12（高表达）
Napsin A	4	2	8（高表达）	4	3	12（高表达）
Ki67	1	3	3（低表达）	2	3	6（高表达）

Fig.3 The detection results of RET fusion genes in parental tissue and its organoid

Fig.4 IC50 results of target drugs

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