Journal of Shanghai Jiao Tong University (Medical Science) >

2024 , Vol. 44 >Issue 4: 435 - 443

DOI: https://doi.org/10.3969/j.issn.1674-8115.2024.04.003

Basic tumor research column

Construction and experimental validation of mouse PDX model by malignant pleural effusion-derived tumor cells from lung cancer

  • WANG Mengting ,
  • CHEN Yinan ,
  • XUANYUAN Xinyang ,
  • YUAN Haihua
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  • Department of Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 201999, China
YUAN Haihua, E-mail: 72300611229@shsmu.edu.cn.

Received date: 2023-10-30

  Accepted date: 2024-03-25

  Online published: 2024-04-28

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Abstract

Objective ·To establish a patient-derived tumor xenograft (PDX) model using tumor cells sourced from malignant pleural effusion (MPE) in patients with lung cancer, and to conduct experimental validation. Methods ·Gene expression data were downloaded from the Gene Expression Omnibus (GEO), including single-cell RNA sequencing data for lung cancer with MPE (GSE131907) and for solid lung cancer (GSE203360). Data were clustered, and differential gene ontology functional enrichment analysis was performed to ascertain the feasibility of modeling by using MPE. MPE samples from patients with lung cancer were collected and processed through centrifugation and red blood cell lysis to enrich cells. The enriched cells were then implanted subcutaneously into non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. Tumor growth was monitored, and when tumors reached 1 000 mm3, they were passaged and preserved. Histopathological examination was conducted on stable passaged tumors, the cell morphology was observed via hematoxylin-eosin (H-E) staining and the expression of lung cancer biomarkers was detected by using immunohistochemistry (IHC). Results ·Single-cell data analysis revealed stronger proliferative functions of tumor cells in MPE, suggesting that PDX modeling using MPE tumor cells may yield better tumor formation. A total of 35 samples of MPE from lung cancer patients were collected, and 13 PDX models were successfully constructed, with a success rate of 37.14%. Histopathological examination showed significant cellular atypia by H-E staining, and IHC result showed positive expression of lung cancer biomarkers such as cytokeratin 7 (CK7), thyroid transcription factor-1 (TTF1), and Napsin A. Conclusion ·By enriching tumor cells from MPE of lung cancer patients, a more convenient, efficient, and dynamically modelable PDX model is successfully constructed. This model retains the malignant characteristics and protein expression features of tumor cells from lung cancer patients, providing an important experimental model tool for basic research and clinical drug guidance for lung cancer patients with MPE.

Key words： lung cancer; malignant pleural effusion (MPE); primary cell culture; patient-derived tumor xenograft (PDX)

Cite this article

WANG Mengting , CHEN Yinan , XUANYUAN Xinyang , YUAN Haihua . Construction and experimental validation of mouse PDX model by malignant pleural effusion-derived tumor cells from lung cancer[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2024 , 44(4) : 435 -443 . DOI: 10.3969/j.issn.1674-8115.2024.04.003

References

1 SIEGEL R L, MILLER K D, FUCHS H E, et al. Cancer statistics, 2022[J]. CA Cancer J Clin, 2022, 72(1): 7-33.
2 ZHENG R S, ZHANG S W, ZENG H M, et al. Cancer incidence and mortality in China, 2016[J]. J Natl Cancer Cent, 2022, 2(1): 1-9.
3 XU P W, TANG K, MA J W, et al. Chemotherapeutic tumor microparticles elicit a neutrophil response targeting malignant pleural effusions[J]. Cancer Immunol Res, 2020, 8(9): 1193-1205.
4 PORCEL J M, GASOL A, BIELSA S, et al. Clinical features and survival of lung cancer patients with pleural effusions[J]. Respirology, 2015, 20(4): 654-659.
5 SHIBAKI R, MURAKAMI S, SHINNO Y, et al. Malignant pleural effusion as a predictor of the efficacy of anti-PD-1 antibody in patients with non-small cell lung cancer[J]. Thorac Cancer, 2019, 10(4): 815-822.
6 SATOH K, MORISAWA S, OKUYAMA M, et al. Severe pleural effusion associated with nilotinib for chronic myeloid leukaemia: cross-intolerance with tyrosine kinase inhibitors[J]. BMJ Case Rep, 2021, 14(9): e243671.
7 WANG T F, CHU S C, LEE J J, et al. Presence of pleural effusion is associated with a poor prognosis in patients with epidermal growth factor receptor-mutated lung cancer receiving tyrosine kinase inhibitors as first-line treatment[J]. Asia Pac J Clin Oncol, 2017, 13(4): 304-313.
8 JIN K T, TENG L S, SHEN Y P, et al. Patient-derived human tumour tissue xenografts in immunodeficient mice: a systematic review[J]. Clin Transl Oncol, 2010, 12(7): 473-480.
9 ROSCILLI G, de VITIS C, FERRARA F F, et al. Human lung adenocarcinoma cell cultures derived from malignant pleural effusions as model system to predict patients chemosensitivity[J]. J Transl Med, 2016, 14: 61.
10 SUN S Q, ZHU J Q, MA Y, et al. Accuracy, robustness and scalability of dimensionality reduction methods for single-cell RNA-seq analysis[J]. Genome Biol, 2019, 20(1): 269.
11 ASHBURNER M, BALL C A, BLAKE J A, et al. Gene ontology: tool for the unification of biology. The Gene Ontology Consortium[J]. Nat Genet, 2000, 25(1): 25-29.
12 WAKEFIELD C E, DOOLAN E L, FARDELL J E, et al. The avatar acceptability study: survivor, parent and community willingness to use patient-derived xenografts to personalize cancer care[J]. EBioMedicine, 2018, 37: 205-213.
13 HEINRICH M A, MOSTAFA A M R H, MORTON J P, et al. Translating complexity and heterogeneity of pancreatic tumor: 3D in vitro to in vivo models[J]. Adv Drug Deliv Rev, 2021, 174: 265-293.
14 MAKIMOTO G, OHASHI K, TOMIDA S, et al. Rapid acquisition of alectinib resistance in ALK-positive lung cancer with high tumor mutation burden[J]. J Thorac Oncol, 2019, 14(11): 2009-2018.
15 PARK B, JEONG B C, CHOI Y L, et al. Development and characterization of a bladder cancer xenograft model using patient-derived tumor tissue[J]. Cancer Sci, 2013, 104(5): 631-638.
16 STOCKHAMMER P, HO C S L, HEGEDUS L, et al. HDAC inhibition synergizes with ALK inhibitors to overcome resistance in a novel ALK mutated lung adenocarcinoma model[J]. Lung Cancer, 2020, 144: 20-29.
17 杨晓乐. 肺癌胸水肿瘤细胞的体外培养、鉴定及其对常规化疗药物敏感性研究[D]. 郑州: 郑州大学, 2017.
17 YANG X L. Establishment and identification of malignant pleural effusion-derived lung cancer primary cell culture and its chemosensitivity[D]. Zhengzhou: Zhengzhou University, 2017.
18 RUAN H Y, WANG Z, SUN Z W, et al. Single-cell RNA sequencing reveals the characteristics of cerebrospinal fluid tumour environment in breast cancer and lung cancer leptomeningeal metastases[J]. Clin Transl Med, 2022, 12(6): e885.
19 KIM N, KIM H K, LEE K, et al. Single-cell RNA sequencing demonstrates the molecular and cellular reprogramming of metastatic lung adenocarcinoma[J]. Nat Commun, 2020, 11(1): 2285.
20 KAO T W, BAI G H, WANG T L, et al. Novel cancer treatment paradigm targeting hypoxia-induced factor in conjunction with current therapies to overcome resistance[J]. J Exp Clin Cancer Res, 2023, 42(1): 171.
21 KANG H N, CHOI J W, SHIM H S, et al. Establishment of a platform of non-small-cell lung cancer patient-derived xenografts with clinical and genomic annotation[J]. Lung Cancer, 2018, 124: 168-178.
22 CHEN X M, SHEN C, WEI Z, et al. Patient-derived non-small cell lung cancer xenograft mirrors complex tumor heterogeneity[J]. Cancer Biol Med, 2021, 18(1): 184-198.
23 CHEN Y, ZHANG R, WANG L, et al. Tumor characteristics associated with engraftment of patient-derived non-small cell lung cancer xenografts in immunocompromised mice[J]. Cancer, 2019, 125(21): 3738-3748.
24 WANG D, PHAM N A, TONG J F, et al. Molecular heterogeneity of non-small cell lung carcinoma patient-derived xenografts closely reflect their primary tumors[J]. Int J Cancer, 2017, 140(3): 662-673.
25 KITA K, FUKUDA K, TAKAHASHI H, et al. Patient-derived xenograft models of non-small cell lung cancer for evaluating targeted drug sensitivity and resistance[J]. Cancer Sci, 2019, 110(10): 3215-3224.
26 STEWART E L, MASCAUX C, PHAM N A, et al. Clinical utility of patient-derived xenografts to determine biomarkers of prognosis and map resistance pathways in EGFR-mutant lung adenocarcinoma[J]. J Clin Oncol, 2015, 33(22): 2472-2480.
27 FICHTNER I, ROLFF J, SOONG R, et al. Establishment of patient-derived non-small cell lung cancer xenografts as models for the identification of predictive biomarkers[J]. Clin Cancer Res, 2008, 14(20): 6456-6468.
28 ZHANG X C, ZHANG J C, LI M, et al. Establishment of patient-derived non-small cell lung cancer xenograft models with genetic aberrations within EGFR, KRAS and FGFR1: useful tools for preclinical studies of targeted therapies[J]. J Transl Med, 2013, 11: 168.
29 LIAO H, ZHOU S X, CHEN S, et al. Establishment and characterization of patient-derived xenograft model of non-small-cell lung cancer derived from malignant pleural effusions[J]. Cancer Manag Res, 2023, 15: 165-174.
30 XIN Y Q, LI S Y, JIANG Q K, et al. Establishment of a jaw fibrosarcoma patient-derived xenograft and evaluation of the tumor suppression efficacy of plumbagin against jaw fibrosarcoma[J]. Front Oncol, 2020, 10: 1479.
31 HUANG T L, CHEN B Y, WANG F, et al. Rab1A promotes IL-4R/JAK1/STAT6-dependent metastasis and determines JAK1 inhibitor sensitivity in non-small cell lung cancer[J]. Cancer Lett, 2021, 523: 182-194.
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