Supplementary Figures S1-S7 from Phage Display-Derived Peptide-Based Dual-Modality Imaging Probe for Bladder Cancer Diagnosis and Resection Postinstillation: A Preclinical Study

Peng, Li; Shang, Wenting; Guo, Pengyu; He, Kunshan; Wang, Hongzhi; Han, Ziyu; Jiang, Hongmei; Tian, Jie; Wang, Kun; Xu, Wanhai

doi:10.1158/1535-7163.22505307

Supplementary Figures S1-S7 from Phage Display-Derived Peptide-Based Dual-Modality Imaging Probe for Bladder Cancer Diagnosis and Resection Postinstillation: A Preclinical Study

journal contribution

posted on 2024-11-04, 08:00 authored by Li Peng, Wenting Shang, Pengyu Guo, Kunshan He, Hongzhi Wang, Ziyu Han, Hongmei Jiang, Jie Tian, Kun Wang, Wanhai Xu

Supplementary Figure S1 shows synthesis and characterization of PLSWT7-DMI. (A) Visible absorption spectrum of PLSWT7-DMI. Similar absorption peaks were observed for PLSWT7-DMI and IRDye800CW (~774 nm). (B) Absorbance stability of PLSWT7-DMI. The absorbance dropped to <10% within 48 h. (C) CCK8 assay of the effects of various concentrations of PLSWT7-DMI on RT112 and EJ cell viability at 24 or 48 h. There were no significant differences in cytotoxicity for all concentrations among groups (p > 0.05). (D) Fluorescence intensity of RT112 and EJ cells incubated with various concentrations of PLSWT7-DMI. Cells incubated with 180 pmol/mL PLSWT7-DMI produced the highest fluorescence. Supplementary Figure S2 shows fluorescence microscopy images of SV-HUC-1 cells incubated with PLSWT7-DMI. Minimal fluorescence was observed after incubation. Supplementary Figure S3 shows metabolic profiles of PLSWT7-DMI post-intravenous-injection via the tail vein. (A) IVIS imaging of subcutaneous tumor-bearing mice injected with cPLSWT7-DMI and PLSWT7-DMI. Red circle indicates the tumor region. (B) Fluorescence intensity of the tumor site in (A). PLSWT7-DMI-treated groups exhibited higher fluorescence intensity than that of cPLSWT7-DMI-treated groups. (C) Comparison of TBR profiles of PLSWT7-DMI and cPLSWT7-DMI. TBR of PLSWT7-DMI-treated groups peaked 8 h post-injection and was higher than the TBR of control groups. Values are presented as means {plus minus} SD of 5 animals. Supplementary Figure S4 shows ex vivo imaging of mouse organs at 6-h post-instillation of PLSWT7-DMI. (A) A bright signal was observed in RT112 tumors, whereas signals in organs were minimal. (B) Quantitative analysis of the fluorescence intensity observed in (A). Values are presented as means {plus minus} SD of 5 animals. Supplementary Figure S5 shows orthotopic BC mice imaging. (A) Luciferase imaging showing tumor growth in the orthotopic BC model. (B) NIR imaging of orthotopic BC mice post-instillation of the probes. Bladders of orthotopic BC mice instilled with PBS, cPLSWT7-DMI, PLSWT7-DMI with blockade, or PLSWT7-DMII, followed by imaging using a homemade NIR-imaging system at 30-min post-instillation. Tumors in PLSWT7-DMI groups were clearly visualized; however, the blockade of PLSWT7-DMI binding to the tumor was observed following PLSWT7 pretreatment. The yellow-dashed circle indicates the tumor site. (C) Quantitative analysis of the mean fluorescence density shown in (B). Values are presented as means {plus minus} SD of 5 animals. *p < 0.05 vs. PLSWT7-DMI+blockade groups. Supplementary Figure S6 shows PAI of subcutaneous tumor-bearing mice after intravenous injection via the tail vein. (A) PAI of subcutaneous tumor-bearing mice. Subcutaneous tumor-bearing mice were injected with PBS, cPLSWT7-DMI, PLSWT7-DMI with blockade, or PLSWT7-DMI via tail vein, followed by PAI. PLSWT7-DMI-treated groups exhibited enhanced contrast compared with other groups. The red-dashed circle indicates the tumor site. (B) Quantification of the average PA-signal amplitude of subcutaneous tumor-bearing mice in (A). The PLSWT7-DMI groups exhibited higher signals than those of other groups. Values present means {plus minus} SD of 5 animals. *p < 0.05 vs. PLSWT7-DMI + blockade groups. Supplementary Figure S7 shows ex vivo human bladder imaging post-administration of cPLSWT7-DMI. No optical contrast between cancer and normal tissues was observed post-administration of cPLSWT7-DMI.

Funding

National Natural Science Foundation of China

National Key R&D Program of China

Chinese Academy of Sciences

Beijing Municipal Science & Technology Commission

Natural Science Foundation of Heilongjiang Province of China

History

ARTICLE ABSTRACT

Bladder cancer is a common human malignancy. Conventional ultrasound and white-light cystoscopy are often used for bladder cancer diagnosis and resection, but insufficient specificity results in a high bladder cancer recurrence rate. New strategies for the diagnosis and resection of bladder cancer are needed. In this study, we developed a highly specific peptide-based probe for bladder cancer photoacoustic imaging (PAI) diagnosis and near-infrared (NIR)-imaging-guided resection after instillation. A bladder cancer–specific peptide (PLSWT7) was selected by in vivo phage-display technology and labeled with IRDye800CW to synthesize a bladder cancer–specific dual-modality imaging (DMI) probe (PLSWT7-DMI). The feasibility of PLSWT7-DMI–based dual-modality PAI-NIR imaging was assessed in vitro, in mouse models, and ex vivo human bladders. An air-pouch bladder cancer (APBC) model suitable for probe instillation was established to evaluate the probe-based bladder cancer PAI diagnosis and NIR-imaging–guided resection. Human bladders were used to assess whether the PLSWT7-DMI–based DMI strategy is a translatable approach for bladder cancer detection and resection. The probe exhibited excellent selectivity and specificity both in vitro and in vivo. Postinstillation of the probe, tumors <3 mm were detectable by PAI, and NIR-imaging–guided tumor resection decreased the bladder cancer recurrence rate by 90% and increased the survival in the mouse model. Additionally, ex vivo NIR imaging of human bladders indicated that PLSWT7-DMI–based imaging would potentially allow precise resection of bladder cancer in clinical settings. This PLSWT7-DMI–based DMI strategy was a translatable approach for bladder cancer diagnosis and resection and could potentially lower the bladder cancer recurrence rate. Mol Cancer Ther; 17(10); 2100–11. ©2018 AACR.