CDH17 nanobodies facilitate rapid imaging of gastric cancer and efficient delivery of immunotoxin

Jingbo Ma; Xiaolong Xu; Chunjin Fu; Peng Xia; Ming Tian; Liuhai Zheng; Kun Chen; Xiaolian Liu; Yilei Li; Le Yu; Qinchang Zhu; Yangyang Yu; Rongrong Fan; Haibo Jiang; Zhifen Li; Chuanbin Yang; Chengchao Xu; Ying Long; Jigang Wang; Zhijie Li

doi:10.1186/s40824-022-00312-3

CDH17 nanobodies facilitate rapid imaging of gastric cancer and efficient delivery of immunotoxin

Biomater Res. 2022 Nov 26;26(1):64. doi: 10.1186/s40824-022-00312-3.

Authors

Jingbo Ma^#^{1

2}, Xiaolong Xu^#³, Chunjin Fu^#³, Peng Xia^{1

4}, Ming Tian^{1

4}, Liuhai Zheng¹, Kun Chen¹, Xiaolian Liu⁵, Yilei Li⁵, Le Yu^{5

6}, Qinchang Zhu⁷, Yangyang Yu⁸, Rongrong Fan⁹, Haibo Jiang¹⁰, Zhifen Li¹¹, Chuanbin Yang³, Chengchao Xu³, Ying Long¹², Jigang Wang^{13

14

15}, Zhijie Li^{16

17}

Affiliations

¹ Department of Hyperbaric Medicine, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, 518020, Guangdong, P. R. China.
² College of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, P. R. China.
³ Department of Geriatrics, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, 518020, Guangdong, P. R. China.
⁴ Department of Hepatobiliary & Pancreatic Surgery, Zhongnan Hospital of Wuhan University, Wuhan, 430072, Hubei, P. R. China.
⁵ Clinical Pharmacy Center, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, P. R. China.
⁶ Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, Guangdong, P.R. China.
⁷ College of Pharmacy, Shenzhen Technology University, Shenzhen, 518118, P.R. China.
⁸ Health Science Center, Shenzhen University, Shenzhen, 518055, Guangdong, P. R. China.
⁹ Deapartment of Biosciences and Nutrition, Karolinska Institute, 14157, Stockholm, Sweden.
¹⁰ Department of Chemistry, The University of Hong Kong, Pok Fu Lam, Hong Kong, P. R. China.
¹¹ School of Chemistry and Chemical Engineering, Shanxi Datong University, Xing Yun Street, Pingcheng District, Datong, 037009, Shanxi, P. R. China.
¹² Department of Hyperbaric Medicine, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, 518020, Guangdong, P. R. China. 13823730482@139.com.
¹³ Department of Geriatrics, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, 518020, Guangdong, P. R. China. jgwang@icmm.ac.cn.
¹⁴ Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, Guangdong, P.R. China. jgwang@icmm.ac.cn.
¹⁵ Artemisinin Research Center, and Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, P. R. China. jgwang@icmm.ac.cn.
¹⁶ Department of Hyperbaric Medicine, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, 518020, Guangdong, P. R. China. li.zhijie@szhospital.com.
¹⁷ Department of Geriatrics, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University, The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen, 518020, Guangdong, P. R. China. li.zhijie@szhospital.com.

^# Contributed equally.

Abstract

Background: It is highly desirable to develop new therapeutic strategies for gastric cancer given the low survival rate despite improvement in the past decades. Cadherin 17 (CDH17) is a membrane protein highly expressed in cancers of digestive system. Nanobody represents a novel antibody format for cancer targeted imaging and drug delivery. Nanobody targeting CHD17 as an imaging probe and a delivery vehicle of toxin remains to be explored for its theragnostic potential in gastric cancer.

Methods: Naïve nanobody phage library was screened against CDH17 Domain 1-3 and identified nanobodies were extensively characterized with various assays. Nanobodies labeled with imaging probe were tested in vitro and in vivo for gastric cancer detection. A CDH17 Nanobody fused with toxin PE38 was evaluated for gastric cancer inhibition in vitro and in vivo.

Results: Two nanobodies (A1 and E8) against human CDH17 with high affinity and high specificity were successfully obtained. These nanobodies could specifically bind to CDH17 protein and CDH17-positive gastric cancer cells. E8 nanobody as a lead was extensively determined for tumor imaging and drug delivery. It could efficiently co-localize with CDH17-positive gastric cancer cells in zebrafish embryos and rapidly visualize the tumor mass in mice within 3 h when conjugated with imaging dyes. E8 nanobody fused with toxin PE38 showed excellent anti-tumor effect and remarkably improved the mice survival in cell-derived (CDX) and patient-derived xenograft (PDX) models. The immunotoxin also enhanced the anti-tumor effect of clinical drug 5-Fluorouracil.

Conclusions: The study presents a novel imaging and drug delivery strategy by targeting CDH17. CDH17 nanobody-based immunotoxin is potentially a promising therapeutic modality for clinical translation against gastric cancer.

Keywords: Cadherin-17; Gastric cancer; Immunotoxin; Jingbo Ma, Xiaolong Xu and Chunjin Fu are contribute equally to this work and share the first-authorship.; Nanobody; Targeted therapy.