GSH/HAase/pH Triple-responsive Nanotheranostic Agent for Synergistic Therapy Guided by Tumor-targeting Fluorescence/Photoacoustic Dual-mode Imaging

doi:10.6023/A24040115

Abstract

In this work, a multifunctional nanotheranostic agent FePDA/DOX@HA-CYS-TTDPP was constructed for photo/chemodynamic/chemo synergistic therapy guided by tumor-targeting fluorescence/photoacoustic dual-mode imaging. The diketopyrrolopyrrole (DPP) derivative (TTDPP) with near-infrared (NIR) absorption was coupled with tumor-targeting biomolecule hyaluronic acid (HA) via cystamine (CYS) to obtain the amphiphilic polymer HA-CYS-TTDPP. It was then self-assembled with Fe³⁺-coordinated polydopamine nanoparticles (FePDA) and anti-tumor drug doxorubicin (DOX) to form this nanomaterial. Its photothermal conversion efficiency reached 65.6% and it showed good photoacoustic performance, which can be attributed to the excellent photothermal conversion performance of TTDPP and PDA. TTDPP was a D-π-A-π-D type DPP derivative formed by the electron acceptor (A) DPP, the electron donor (D) triphenylamine, and the conjugated bridge thiophene, which exhibited excellent photothermal and photoacoustic performance, and a certain degree of photodynamic effect. PDA was formed by dopamine oxidation self-polymerization, which showed the advantages of good biocompatibility, high photothermal conversion efficiency, and easy functionalization; moreover, due to the d-d transition of exogenous Fe³⁺in FePDA, the photothermal effect of PDA can be further enhanced. Furthermore, the weak acidity of tumor microenvironment (TME) facilitated the dissolution and release of DOX, and the overexpressed hyaluronidase (HAase) and glutathione (GSH) in tumor cells can also trigger the degradation of HA and the cleavage of disulfide bonds in CYS, respectively, leading to the release of DOX. Thus, the distance between DOX and PDA and TTDPP was increased, and the fluorescence of DOX was recovered, thereby achieving TME-responsive fluorescence activation and controlled drug release, and the tumor-targeting effect was further enhanced. Therefore, this nanotheranostic agent can realize triple response to the weak acidity, overexpressed GSH and HAase of TME. On the other hand, Fe³⁺ can react with GSH and H₂O₂ in tumor cells to induce Fenton reaction, generating cytotoxic •OH and resulting in chemodynamic therapeutic performance. Cell experiments confirmed that FePDA/DOX@HA-CYS-TTDPP can intelligently respond to TME and achieve targeted fluorescence imaging, and effectively inhibit the proliferation of HeLa tumor cells through photo/chemodynamic/chemo synergistic therapy. This system provides new ideas for achieving precise and efficient tumor theranostics.

Key words: tumor microenvironment-responsive, dual-mode imaging, photothermal therapy, chemodynamic therapy, synergistic therapy

Cite this article

Yanqin Huang, Jiwen Luo, Jiaqi Li, Rui Zhang, Xingfen Liu, Quli Fan, Wei Huang. GSH/HAase/pH Triple-responsive Nanotheranostic Agent for Synergistic Therapy Guided by Tumor-targeting Fluorescence/Photoacoustic Dual-mode Imaging[J]. Acta Chimica Sinica, 2024, 82(8): 903-913.

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Fig. & Tab. 10

Figure 1. (a) Synthetic routes of HA-CYS-TTDPP; (b) schematic illustration of the preparation of FePDA/DOX@HA-CYS-TTDPP and the tumor- targeting FLI/PAI-guided synergistic PTT/PDT/CDT/CHT

Figure 2. (a) UV-Vis absorption spectra of TTDPP and HA-CYS-TTDPP in DMF, and FePDA@HA-CYS-TTDPP, FePDA/DOX@HA-CYS-TTDPP, FePDA and DOX in H2O; (b) TEM image of FePDA (Insert of b: average hydrodynamic sizes); (c, d) TEM images of FePDA/DOX@HA-CYS-TTDPP (Inset of c: average hydrodynamic sizes)

Figure 3. Emission spectra of FePDA/DOX@HA-CYS-TTDPP at 37 ℃ with excitation at 480 nm in different conditions: (a) during 24 h treatment with DTT (10 mmol/L) in PBS (0.02 mol/L, pH 7.4); (b) before and after treatment with HAase (10 U/mL) in PBS (0.02 mol/L, pH 5.6) for 1 h

Figure 4. Average hydrodynamic sizes of FePDA/DOX@HA-CYS-TTDPP at 37 ℃ in different conditions: (a) after treatment with DTT (10 mmol/L) in PBS (0.02 mol/L, pH 7.4) for 4 h (inset: TEM image under this condition); (b) after treatment with HAase (10 U/mL) in PBS (0.02 mol/L, pH 5.6) for 1 h (inset: TEM image under this condition)

Figure 5. In a NaHCO3 buffer solution (25 mmol/L) containing GSH (10 μg/mL), H2O2 (10 mmol/L) and MB (5 mmol/L), (a) absorbance changes of MB over time after adding FePDA/DOX@HA-CYS-TTDPP; (b) absorbance changes of MB over time without FePDA/DOX@HA-CYS-TTDPP

Figure 6. (a) Temperature elevation curves FePDA/DOX@HA-CYS-TTDPP (0.16 mg/mL) under laser irradiation with different powers (635 nm, 0.8~1.2 W/cm2); (b) photothermal response of FePDA/DOX@HA-CYS-TTDPP under laser irradiation (635 nm, 1 W/cm2) with repeated irradiation and shutdown cycle five times; (c) single photothermal cycle of the process in (b); (d) linear time versus －lnθ obtained from the cooling period of (c)

Figure 7. (a) UV-Vis absorption spectra of DPBF in the aqueous solutions containing FePDA/DOX@HA-CYS-TTDPP at different irradiation time (635 nm, 30 mW/cm2); (b) photodegradation rate of DPBF in the aqueous solution containing MB or FePDA/DOX@HA-CYS-TTDPP; (c) 1O2 detection in HeLa cells after incubation with FePDA/DOX@HA-CYS-TTDPP, using DCFH-DA as a probe (635 nm, 100 mW/cm2)

Figure 8. Photoacoustic signals of the aqueous solutions of FePDA/ DOX@HA-CYS-TTDPP with different concentrations (inset: photoacoustic images corresponding to different concentrations)

Figure 9. (a) NIR fluorescence images of (1) HeLa cells, (2) HeLa cells pretreated with HA, and (3) NIH-3T3 cells after incubation with FePDA/DOX@HA-CYS-TTDPP for 4 h; (b) changes in fluorescence intensity over time in HeLa cells incubated with FePDA/DOX@HA-CYS-TTDPP for 30 min (the excitation wavelength is 480 nm)

Figure 10. (a) Cell viability of NIH-3T3 cells treated with FePDA/DOX@HA-CYS-TTDPP under dark conditions; (b) cell viability of HeLa cells treated with FePDA/DOX@HA-CYS-TTDPP without or with irradiation (635 nm, 1 W/cm2) for 5 min; (c) AM/PI staining of HeLa cells treated with FePDA/DOX@HA-CYS-TTDPP without or with irradiation (635 nm, 1 W/cm2) for 10 min

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GSH/HAase/pH三重响应纳米诊疗剂用于肿瘤靶向荧光/光声双模成像指导的协同治疗