TMEM16A-inhibitor loaded pH-responsive nanoparticles: A novel dual-targeting antitumor therapy for lung adenocarcinoma
Graphical abstract
Introduction
Lung cancer is the most commonly diagnosed malignancy and is associated with the highest mortality rate [1], [2]. Non-small-cell lung cancer (NSCLC) accounts for more than 85% of lung cancer, and 54% of NSCLC is lung adenocarcinoma [3], [4]. Currently, the main method for treating advanced lung adenocarcinoma is chemotherapy [5]. Since chemotherapy drugs are accompanied with serious side effects, it is urgent to identify novel drug targets and develop safe and efficient antitumor strategy [6].
As nanocarriers possess desirable properties of targeting, compatibility and sustained release ability, the nanocarriers delivery systems have been widely explored for tumor therapy exploration. Accurate targeting of tumor tissue are highly desirable to improve based on current nanocarriers because it will greatly reduce the side effects during the cancer targeting process. For enhancing the targeting of anti-tumor drugs, such as doxorubicin, paclitaxel and cisplatin, adopting nanocarriers is an important trend in the development of cancer therapy [7], [8], [9]. However, the current nano-delivery systems can not completely eliminate the high probability of kidney toxicity, neurotoxicity and toxicity to the hematopoietic system and digestive system of the chemotherapy drugs [10], [11]. Therefore, improving the properties of nanocarriers have become urgent issues [12], [13].
The identification of novel and tumor-specific drug targets is another main focus of drug development. Recently, molecular targeted therapy for treating lung adenocarcinoma has shown some promising results, like epidermal growth factor tyrosine kinase inhibitors (EGFR-TKIs). However, these TKIs also produce drug resistance, diarrhea, dermatitis, stomatitis and other unwanted side effects [14], [15], [16]. More and more studies have shown that ion channels may be important drug targets for the treatment of lung adenocarcinoma [17], [18]. Our previous studies have shown that TMEM16A, which is highly expressed in lung adenocarcinoma cells, is one of the key targets for treating lung adenocarcinoma [19]. Compared with existing cytotoxic antitumor drugs, the antitumor drugs targeting ion channels have little resistance and toxic side effects. TMEM16A was highly overexpressed in the lung adenocarcinoma cell line LA795 [19]. The Tmem16a gene is located in the human chromosome band 11q13, which is one of the most frequently amplified regions in human cancer [20], [21]. TMEM16A is highly expressed in a variety of cancer cells, therefore TMEM16A is considered as a diagonosis cancer marker and antitumor drug target [22], [23].
In order to improve the anti-cancer efficiency, a targeted nanocarrier system and a TMEM16A inhibitor were combined to develop a novel therapy against lung adenocarcinoma. In this work, we designed a novel pH-sensitive nanocarrier, PEO-b-P(DMAEMA-co-MAEBA), that can specifically deliver drugs to tumor tissues because of the weak acidity microenvironment of lung adenocarcinoma. The nanocarrier can be accumulated in the tumor lesions, thereby increasing the local drug concentration to enhance efficacy. We also tested a previously identified specific inhibitor of TMEM16A, CaCCinh-A01 (A01), and found that it can suppress cancer cell growth by inhibiting TMEM16A. By combining the specific inhibitor of TMEM16A with the pH-sensitive nanocarrier, we developed a dual-targeting anti-cancer system that targets cancer tissue and TMEM16A ion channel, respectively. The results showed this dual-targeting antitumor strategy improved the therapeutic effect to more than two times compared with the therapeutic effect of direct A01 administration, and this new system has no observable side effects.
Section snippets
Materials
A01 was purchased from MedChemExpress (CAS No.: 407587-33-1; New Jersey, USA). DiI (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate, CAS No.: 41085-99-8) and DMSO (dimethyl sulfoxide, CAS No.: 67–68-5) were purchased from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). The reagents 4% paraformaldehyde, goat serum, RPMI-1640 medium and MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide, CAS No.: 298-93-1) were purchased from Solarbio (Beijing,
A01 inhibited the TMEM16A current in LA795 cells in a concentration-dependent manner.
We found that in immunofluorescence experiments the endogenous TMEM16A protein was highly expressed in LA795 cells (Fig. 1A). It has been reported that A01 is a specific inhibitor of TMEM16A [28]. For validation, we performed whole-cell patch clamp recordings on HEK293 cells in which TMEM16A was transiently transfected. TMEM16A is a Ca2+-activated Cl- channel [29] and whole-cell recordings were achieved by pipette solutions containing 600 nM Ca2+. Similar to previous reports [30], these
Discussion
In this study, we designed a dual-targeting antitumor strategy via the combination of pH-sensitive copolymers (PEO-b-P(DMAEMA-co-MAEBA)), which can specifically deliver drugs to cancer cells, with a specific inhibitor (A01) of TMEM16A, which is overexpressed in lung adenocarcinoma cells (Fig. 9). Patch clamp results showed that A01 inhibited the TMEM16A currents in LA795 cells in a concentration-dependent manner with an IC50 of 7.35 ± 0.86 μM. Then, we demonstrated that A01 could inhibit the
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
This work was supported by the National Natural Science Foundation of China (Grant No. 11735006 to Y Zhan, 81830061 to HL An), the Natural Science Foundation of Tianjin of China (Grant No. 19JCYBJC28300 to HL An), the Natural Science Foundation of Hebei Province of China (Grant No. C2018202302 to YF Chen), the Youth Talent Support Program of Hebei Province of China (Grant No.2013001 to YF Chen).
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2024, International Journal of Biological MacromoleculesTMEM16A ion channel: A novel target for cancer treatment
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2022, International Journal of Biological MacromoleculesCitation Excerpt :In addition, TMEM16A also forms an interaction network with EGFR proteins and ezrin–radixin–moesin (ERM) proteins, and it regulates the growth of cancer cells through signal transduction [34,35]. TMEM16A is only expressed in specific normal tissues but is abnormally upregulated in cancer cells, so it is a suitable drug target for lung adenocarcinoma treatment [36]. In this work, virtual screening found that the binding affinity of natural product narirutin and TMEM16A protein was high.
Self-healing pectin/cellulose hydrogel loaded with limonin as TMEM16A inhibitor for lung adenocarcinoma treatment
2022, International Journal of Biological MacromoleculesCitation Excerpt :The TMEM16A as a calcium-activated chloride channel [6] is expressed in oral salivary glands, respiratory tract, alveoli, pancreas and other fluid-secreting organs normally [7,8], but specifically over expressed in several tumor tissues such as lung cancer, head and neck squamous carcinoma, liver cancer, and colorectal cancer [9–11]. High expression of TMEM16A promoted the proliferation and migration of these cancer cells, and lead to a poor prognosis [12], while inhibiting of TMEM16A significantly inhibited the development of these cancers [13]. Therefore, TMEM16A is considered as a new drug target for cancer treatment and natural compounds as TMEM16A inhibitors were expected to be screened and effectively inhibit the growth of lung adenocarcinoma.
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2022, Biomedicine and PharmacotherapyCitation Excerpt :TMEM16A overexpression is closely related to cancer growth, metastasis, and prognosis [19]. The proliferation and migration of cancer cells were suppressed, and cells apoptosis were induced by TMEM16A inhibition [20,21]. Therefore, screen of anticancer drugs by regulating TMEM16A is a feasible therapeutic option.
Molecular mechanism of CaCC<inf>inh</inf>-A01 inhibiting TMEM16A channel
2020, Archives of Biochemistry and BiophysicsCitation Excerpt :A piece of encouraging news, the development of drug delivery systems solved this problem very well. We recently developed a strategy to precisely deliver A01 to the tumor region using pH-responsive nanoparticles, which resolved the effects of the drug on normal tissues [51]. Our results have made progress in revealing the molecular mechanism of A01 inhibition of TRMEM16A, and these results will facilitate the discovery and design of novel TMEM16A inhibitors in the future.
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These authors contributed equally to this work.