Application of high-throughput drug sensitivity screening system in the treatment of non-small cell lung cancer with malignant pleural effusion

doi:10.3969/j.issn.1674-8115.2022.01.012

Abstract

Abstract: Objective

·To explore the feasibility of individualized treatment of malignant pleural effusion (MPE) patients with non-small cell lung cancer (NSCLC) guided by in vitro high-throughput drug sensitivity screening system.

Methods

·Thirty NSCLC patients with MPE treated in Nanjing Chest Hospital from January 2019 to May 2020 were included. The tumor cells in MPE were collected and isolated, and primary cells were cultured. The ex vivo drug sensitivity of primary cells was assessed for 30 chemotherapeutical and molecular targeted drugs by using the high-throughput drug sensitivity screening system of automated digital fluorescence imager. The consistency between the drug sensitivity assay results and the clinical efficacy of patients was evaluated.

Results

·The ex vivo drug sensitivity results of primary tumor cells cultured from MPE revealed considerable heterogeneity in drug response. Most of the primary cells were sensitive to Epirubicin, Lobaplatin, Gemcitabine and Irinotecan. Clinical efficacy of 23 patients can be evaluated. For patients with previously untreated, epidermal growth factor receptor (EGFR) mutation-positive advanced NSCLC, the consistency between the efficacy of targeted therapy and the high-throughput drug sensitivity assay results was 44.4% (4/9). For the EGFR negative untreated patients and EGFR mutation-positive advanced NSCLC patients with acquired resistance to targeted treatment, the consistency between the therapeutic effect of chemotherapy and the high-throughput drug sensitivity assay results was 85.7% (12/14).

Conclusion

·The high-throughput drug sensitivity assay based on MPE primary cell culture in NSCLC patients has a high consistency with the clinical efficacy of chemotherapy, which is helpful for decision-making of personalized chemotherapy.

Key words: non-small cell lung cancer, pleural effusion, in vitro chemosensitivity assay, individualized treatment

CLC Number:

R734.2

Yu ZHANG, Xiaoyuan WU, Lihua GUAN, Yiyuan LIU, Xingyue PENG, Haiyan XIE, Wei HU, Keke HAO, Ning XIA, Guojun LU, Zhibo HOU. Application of high-throughput drug sensitivity screening system in the treatment of non-small cell lung cancer with malignant pleural effusion[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2022, 42(1): 82-89.

Figures/Tables 5

TrendMD

References 30

1	BRAY F, FERLAY J, SOERJOMATARAM I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68(6): 394-424.
2	SIEGEL R L, MILLER K D, JEMAL A. Cancer statistics, 2018[J]. CA Cancer J Clin, 2018, 68(1): 7-30.
3	BURROWS C M, MATHEWS W C, COLT H G. Predicting survival in patients with recurrent symptomatic malignant pleural effusions: an assessment of the prognostic values of physiologic, morphologic, and quality of life measures of extent of disease[J]. Chest, 2000, 117(1): 73-78.
4	CUFER T, KNEZ L. Update on systemic therapy of advanced non-small-cell lung cancer[J]. Expert Rev Anticancer Ther, 2014, 14(10): 1189-1203.
5	SZULKIN A, OTVÖS R, HILLERDAL C O, et al. Characterization and drug sensitivity profiling of primary malignant mesothelioma cells from pleural effusions[J]. BMC Cancer, 2014, 14: 709.
6	ÖTVÖS R, SZULKIN A, HILLERDAL C O, et al. Drug sensitivity profiling and molecular characteristics of cells from pleural effusions of patients with lung adenocarcinoma[J]. Genes Cancer, 2015, 6(3/4): 119-128.
7	中国临床肿瘤学会指南工作委员会组织. 中国临床肿瘤学会(CSCO)原发性肺癌诊疗指南-2019[M]. 北京: 人民卫生出版社, 2019.
8	LEE S H. Chemotherapy for lung cancer in the era of personalized medicine[J]. Tuberc Respir Dis (Seoul), 2019, 82(3): 179-189.
9	FRIBOULET L, OLAUSSEN K A, PIGNON J P, et al. ERCC1 isoform expression and DNA repair in non-small-cell lung cancer[J]. N Engl J Med, 2013, 368(12): 1101-1110.
10	OLAUSSEN K A, POSTEL-VINAY S. Predictors of chemotherapy efficacy in non-small-cell lung cancer: a challenging landscape[J]. Ann Oncol, 2016, 27(11): 2004-2016.
11	SEUFFERLEIN T, AHN J, KRNDIJA D, et al. Tumor biology and cancer therapy: an evolving relationship[J]. Cell Commun Signal, 2009, 7: 19.
12	BEN-DAVID U, SIRANOSIAN B, HA G, et al. Genetic and transcriptional evolution alters cancer cell line drug response[J]. Nature, 2018, 560(7718): 325-330.
13	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.
14	FULCHER M L, RANDELL S H. Human nasal and tracheo-bronchial respiratory epithelial cell culture[J]. Methods Mol Biol, 2013, 945: 109-121.
15	WILDING J L, BODMER W F. Cancer cell lines for drug discovery and development[J]. Cancer Res, 2014, 74(9): 2377-2384.
16	BYRNE A T, ALFÉREZ D G, AMANT F, et al. Interrogating open issues in cancer precision medicine with patient-derived xenografts[J]. Nat Rev Cancer, 2017, 17(4): 254-268.
17	GAO H, KORN J M, FERRETTI S, et al. High-throughput screening using patient-derived tumor xenografts to predict clinical trial drug response[J]. Nat Med, 2015, 21(11): 1318-1325.
18	ROSSI G, MANFRIN A, LUTOLF M P. Progress and potential in organoid research[J]. Nat Rev Genet, 2018, 19(11): 671-687.
19	DE WETERING MVAN, FRANCIES H E, FRANCIS J M, et al. Prospective derivation of a living organoid biobank of colorectal cancer patients[J]. Cell, 2015, 161(4): 933-945.
20	CUNDERLÍKOVÁ B. Issues to be considered when studying cancer in vitro[J]. Crit Rev Oncol Hematol, 2013, 85(2): 95-111.
21	YIN S Y, XI R B, WU A W, et al. Patient-derived tumor-like cell clusters for drug testing in cancer therapy[J]. Sci Transl Med, 2020, 12(549): eaaz1723.
22	MARKASZ L, KIS L L, STUBER G, et al. Hodgkin-lymphoma-derived cells show high sensitivity to dactinomycin and paclitaxel[J]. Leuk Lymphoma, 2007, 48(9): 1835-1845.
23	SKRIBEK H, OTVOS R, FLABERG E, et al. Chronic lymphoid leukemia cells are highly sensitive to the combination of prednisolone and daunorubicin, but much less to doxorubicin or epirubicin[J]. Exp Hematol, 2010, 38(12): 1219-1230.
24	DEDINSZKI D, KISS A, MÁRKÁSZ L, et al. Inhibition of protein phosphatase-1 and-2A decreases the chemosensitivity of leukemic cells to chemotherapeutic drugs[J]. Cell Signal, 2015, 27(2): 363-372.
25	MANCINI R, GIARNIERI E, DE VITIS C, et al. Spheres derived from lung adenocarcinoma pleural effusions: molecular characterization and tumor engraftment[J]. PLoS One, 2011, 6(7): e21320.
26	SZULKIN A, NILSONNE G, MUNDT F, et al. Variation in drug sensitivity of malignant mesothelioma cell lines with substantial effects of selenite and bortezomib, highlights need for individualized therapy[J]. PLoS One, 2013, 8(6): e65903.
27	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.
28	ALIZADEH A A, ARANDA V, BARDELLI A, et al. Toward understanding and exploiting tumor heterogeneity[J]. Nat Med, 2015, 21(8): 846-853.
29	王丹. 高通量体外药敏检测技术在晚期肺癌伴恶性胸水中的临床研究[D]. 合肥: 安徽医科大学, 2018: 1-41.
30	中华医学会, 中华医学会肿瘤学分会, 中华医学会杂志社. 中华医学会肺癌临床诊疗指南(2018版)[J]. 中华肿瘤杂志, 2018, 40(12): 935-964.

Sample	Gender	Age /year	Therapy line	Histology	EGFR/ALK/ ROS1 testing	Treatment	Clinical efficacy	High-throughput drug sensitivity test in vitro	Consistency analysis
S1	Male	72	1	Adenocarcinoma	EGFR19DEL	Gefitinib	PR	Gefitinib resistance	Inconsistent
S3	Female	48	1	Adenocarcinoma	EGFR19DEL	Gefitinib	PR	Gefitinib resistance	Inconsistent
S4	Female	78	1	Adenocarcinoma	Negative	Pemetrexed + Carboplatin	SD	Low sensitive to Pemetrexed and Carboplatin	Consistent
S5	Female	48	1	Adenocarcinoma	EGFR19DEL	Gefitinib	PR	Low sensitive to Gefitinib	Consistent
S7	Female	48	1	Adenocarcinoma	Negative	Pemetrexed + Cisplatin + Bevacizumab	PR	Low sensitive to Pemetrexed and Cisplatin	Consistent
S10	Female	60	1	Adenocarcinoma	Negative	Pemetrexed + Carboplatin	SD	Low sensitive to Pemetrexed and Carboplatin	Consistent
S15	Female	67	1	Adenocarcinoma	Negative	Pemetrexed + Cisplatin	PR	Resistant to Pemetrexed and Cisplatin	Inconsistent
S16	Male	63	1	Adenocarcinoma	EGFR21L858R	Gefitinib	SD	Low sensitive to Gefitinib	Consistent
S19	Male	74	1	Adenosquamous	Negative	Docetaxel + Nedaplatin	SD	Low sensitive to Docetaxel and Nedaplatin	Consistent
S20	Female	57	1	Adenocarcinoma	EGFR19DEL	Afatinib	PR	Afatinib resistance	Inconsistent
S23	Male	65	1	Adenocarcinoma	EGFR19DEL	Gefitinib	PR	Gefitinib resistance	Inconsistent
S24	Male	77	1	Adenocarcinoma	Negative	Pemetrexed + Carboplatin	SD	Resistant to Pemetrexed and Carboplatin	Inconsistent
S25	Male	74	1	Adenocarcinoma	EGFR21L858R	Gefitinib	PR	Moderate sensitive to Gefitinib	Consistent
S26	Female	66	1	Adenocarcinoma	EGFR21L861Q	Afatinib	PR	Low sensitive to Afatinib	Consistent
S27	Male	92	1	Adenocarcinoma	EGFR19DEL	Gefitinib	SD	Gefitinib resistance	Inconsistent
S8	Male	45	3	Adenocarcinoma	Negative	Vinorelbine + Lobaplatin	SD	Low sensitive to Vinorelbine， moderate sensitive to Lobaplatin	Consistent
S11	Female	48	3	Adenocarcinoma	EGFR19DEL	Pemetrexed + Carboplatin + Bevacizumab	PR	Moderate sensitive to Pemetrexed and Carboplatin	Consistent
S13	Male	55	2	Squamous	Negative	Gemcitabine + Nedaplatin	SD	Moderate sensitive to Gemcitabine， low sensitive to Nedaplatin	Consistent
S14	Female	82	3	Adenocarcinoma	EGFR21L858R/T790M/C797S（trans）	S1+Anlotinib	SD	Low sensitive to Tegafur and Nintedanib	Consistent
S18	Male	72	3	Squamous	Negative	S1	SD	Low sensitive to Tegafur	Consistent
S21	Male	45	5	Adenocarcinoma	EGFR19DEL/ T790M/C797S（trans）	Gemcitabine + Lobaplatin	PR	High sensitive to Gemcitabine， moderate sensitive to Lobaplatin	Consistent
S22	Female	71	3	Adenocarcinoma	EGFR21L858R/T790M/C797S（trans）	Pemetrexed + Carboplatin +Bevacizumab	PR	Moderate sensitive to Pemetrexed and Carboplatin	Consistent
S29	Female	60	3	Adenocarcinoma	EGFR21L858R+BRAF V600E	Pemetrexed + Carboplatin	PD	Resistant to Pemetrexed and Carboplatin	Consistent

Sample	Gender	Age /year	Therapy line	Histology	EGFR/ALK/ ROS1 testing	Treatment	Clinical efficacy	High-throughput drug sensitivity test in vitro	Consistency analysis
S1	Male	72	1	Adenocarcinoma	EGFR19DEL	Gefitinib	PR	Gefitinib resistance	Inconsistent
S3	Female	48	1	Adenocarcinoma	EGFR19DEL	Gefitinib	PR	Gefitinib resistance	Inconsistent
S4	Female	78	1	Adenocarcinoma	Negative	Pemetrexed + Carboplatin	SD	Low sensitive to Pemetrexed and Carboplatin	Consistent
S5	Female	48	1	Adenocarcinoma	EGFR19DEL	Gefitinib	PR	Low sensitive to Gefitinib	Consistent
S7	Female	48	1	Adenocarcinoma	Negative	Pemetrexed + Cisplatin + Bevacizumab	PR	Low sensitive to Pemetrexed and Cisplatin	Consistent
S10	Female	60	1	Adenocarcinoma	Negative	Pemetrexed + Carboplatin	SD	Low sensitive to Pemetrexed and Carboplatin	Consistent
S15	Female	67	1	Adenocarcinoma	Negative	Pemetrexed + Cisplatin	PR	Resistant to Pemetrexed and Cisplatin	Inconsistent
S16	Male	63	1	Adenocarcinoma	EGFR21L858R	Gefitinib	SD	Low sensitive to Gefitinib	Consistent
S19	Male	74	1	Adenosquamous	Negative	Docetaxel + Nedaplatin	SD	Low sensitive to Docetaxel and Nedaplatin	Consistent
S20	Female	57	1	Adenocarcinoma	EGFR19DEL	Afatinib	PR	Afatinib resistance	Inconsistent
S23	Male	65	1	Adenocarcinoma	EGFR19DEL	Gefitinib	PR	Gefitinib resistance	Inconsistent
S24	Male	77	1	Adenocarcinoma	Negative	Pemetrexed + Carboplatin	SD	Resistant to Pemetrexed and Carboplatin	Inconsistent
S25	Male	74	1	Adenocarcinoma	EGFR21L858R	Gefitinib	PR	Moderate sensitive to Gefitinib	Consistent
S26	Female	66	1	Adenocarcinoma	EGFR21L861Q	Afatinib	PR	Low sensitive to Afatinib	Consistent
S27	Male	92	1	Adenocarcinoma	EGFR19DEL	Gefitinib	SD	Gefitinib resistance	Inconsistent
S8	Male	45	3	Adenocarcinoma	Negative	Vinorelbine + Lobaplatin	SD	Low sensitive to Vinorelbine， moderate sensitive to Lobaplatin	Consistent
S11	Female	48	3	Adenocarcinoma	EGFR19DEL	Pemetrexed + Carboplatin + Bevacizumab	PR	Moderate sensitive to Pemetrexed and Carboplatin	Consistent
S13	Male	55	2	Squamous	Negative	Gemcitabine + Nedaplatin	SD	Moderate sensitive to Gemcitabine， low sensitive to Nedaplatin	Consistent
S14	Female	82	3	Adenocarcinoma	EGFR21L858R/T790M/C797S（trans）	S1+Anlotinib	SD	Low sensitive to Tegafur and Nintedanib	Consistent
S18	Male	72	3	Squamous	Negative	S1	SD	Low sensitive to Tegafur	Consistent
S21	Male	45	5	Adenocarcinoma	EGFR19DEL/ T790M/C797S（trans）	Gemcitabine + Lobaplatin	PR	High sensitive to Gemcitabine， moderate sensitive to Lobaplatin	Consistent
S22	Female	71	3	Adenocarcinoma	EGFR21L858R/T790M/C797S（trans）	Pemetrexed + Carboplatin +Bevacizumab	PR	Moderate sensitive to Pemetrexed and Carboplatin	Consistent
S29	Female	60	3	Adenocarcinoma	EGFR21L858R+BRAF V600E	Pemetrexed + Carboplatin	PD	Resistant to Pemetrexed and Carboplatin	Consistent

[1]	Yun-fang MA, Li-na PAN, Zhen LI, Bei-li GAO, Jia-an HU, Zhi-hong XU. Exploratory study on downregulation of PD-L1 in KRAS G12V-mutant non-small cell lung cancer cells by selumetinib [J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(6): 741-748.
[2]	QU Guo-jun1, LU Yuan-feng2, LI Yu1. Mechanism of CCT2, a new downstream substrate of PDGFRα, on proliferation of tumor cells [J]. , 2019, 39(1): 28-.
[3]	HE Ya-ping1, LOU Wei-qun2, CHEN Jie-ling2, ZHOU Zhen3, ZHU Jing-fen1, JIAN Hong3. Influencing factors for quality of life in patients over 12 months after diagnosis of non-small cell lung cancer [J]. , 2018, 38(7): 775-.
[4]	CHEN Lei, XIA Shu-yue. Therapeutic effect and safety of bevacizumab combined with cisplatin on malignant pleural effusion of patients with non-small cell lung cancer [J]. , 2015, 35(8): 1194-.
[5]	SHI Qin, CHEN Qun, XIE Qiang, et al. Effects of mRNA expressions of ERCC1, RRM1, TYMS, and TUBB3 of NSCLC patients with wild-type EGFR on efficacy and prognosis of chemotherapy [J]. , 2015, 35(7): 1010-.
[6]	LI Yong, LI Zi-ming, LU Shun. Distribution of EGFR gene mutations of early stage non-small cell lung cancer [J]. , 2014, 34(4): 511-.
[7]	WANG Lin, LOU Jia-tao. Effects of microRNA-128a on proliferation and migration of non-small cell lung cancer cells [J]. , 2014, 34(3): 268-.
[8]	DU Wei, ZHANG Xiao, GAO Cui, et al. New grey correlation analysis of hospitalization expenses of 600 patients with non-small cell lung cancer [J]. , 2012, 32(10): 1356-.
[9]	ZHU Li-min, ZHANG Hui, ZHOU Hao, et al. Time-effect relationship of Kangliu Zengxiao Decoction in reducing chemotherapy-associated adverse reactions in patients with advanced non-small cell lung cancer [J]. , 2011, 31(6): 797-.
[10]	LU Shun. New approaches for individual treatment of advanced non-small cell lung cancer [J]. , 2011, 31(3): 261-.
[11]	LOU Jia-tao, WU Chuan-yong, XUE Jian, et al. Preliminary establishment and application of a new method for detection of EGFR mutations based on suspension array technology [J]. , 2011, 31(3): 284-.
[12]	GAO Zhi-qiang, HAN Bao-hui, SHEN Jie, et al. Expression of ERCC1, RRM1 and BRCA1 in tissues of advanced non-small cell lung cancer [J]. , 2011, 31(3): 290-.
[13]	SONG Yi-yi, GU Ai-qin, HAN Bao-hui. Expression of p21, p53 and c-erbB-2 in tissues of non-small cell lung cancer and its clinical significance [J]. , 2011, 31(3): 295-.
[14]	GU Ai-qin, GAO Zhi-qiang, WANG Hui-min, et al. Clinical analysis of gefitinib in treatment of elderly patients with advanced non-small cell lung cancer [J]. , 2011, 31(3): 305-.
[15]	BAI Hao, HAN Bao-hui. Role of surgical resection in comprehensive treatment for patients with non-small cell lung cancer and solitary brain metastasis [J]. , 2011, 31(3): 309-.