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. 2021 Aug 11;13(8):1239.
doi: 10.3390/pharmaceutics13081239.

Development, Characterization, and In Vivo Evaluation of a Novel Aptamer (Anti-MUC1/Y) for Breast Cancer Therapy

Huma Khan  1 , Vaidehi Makwana  1 , Sofia Nascimento Dos Santos  2 , Carlos Eduardo Bonacossa de Almeida  3 , Ralph Santos-Oliveira  4   5 , Sotiris Missailidis  6
Affiliations

Development, Characterization, and In Vivo Evaluation of a Novel Aptamer (Anti-MUC1/Y) for Breast Cancer Therapy

Huma Khan et al. Pharmaceutics. .

Abstract

MUC1, the transmembrane glycoprotein Mucin 1, is usually found to be overexpressed in a variety of epithelial cancers playing an important role in disease progression. MUC1 isoforms such as MUC1/Y, which lacks the entire variable number of tandem repeat region, are involved in oncogenic processes by enhancing tumour initiation. MUC1/Y is therefore considered a promising target for the identification and treatment of epithelial cancers; but so far, the precise role of MUC1/Y remains to be elucidated. In this work, we developed and identified a DNA aptamer that specifically recognizes the splice variant MUC1/Y for the first time. The DNA aptamer could bind to a wide variety of human cancer cells, and treatment of MUC1/Y positive cells resulted in reduced growth in vitro. Moreover, MUC1/Y aptamer inhibited the tumour growth of breast cancer cells in vivo. The present study highlights the importance of targeting MUC1/Y for cancer treatment and unravels the suitability of a DNA aptamer to act as a new therapeutic tool.

Keywords: MUC1/Y; aptamer; cancer; pharmacokinetics; therapy.

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Conflict of interest statement

The authors declare no conflict of interest concerning this article.

Figures

Figure 1
Figure 1
The three selected aptamers, (A) S11, (B) S51, and (C) S75 in three presentations: entire aptamer (left), variable region (middle), and structured region (right). All dG values are expressed in Kcal/mol.
Figure 2
Figure 2
Fluorescence changes observed upon titration of the 20mer into the S11 aptamer-dye complex. (A) Emission spectra; (B) Fluorescence changes plot as a function of 20mer concentration. All experiments were performed in duplicate and the results presented are the average of these measurements; (C) Dissociation constants obtained from the Ru (II) complex displacement assay. Data for the aptamer-dye complex were all fit to a sigmoidal function while data for the aptamer-peptide complex best fit to differing functions: MM = standard hyperbolic function described by the Michaelis–Menten equation; SF = sigmoidal fit; and QD = quadratic function for binding.
Figure 3
Figure 3
Thermal denaturation of S11a. (A) Fluorescence melting profiles of S11a alone at 3 TDC; (B) Fluorescence melting profiles of S11a with the 20mer peptide at 3 TDC.
Figure 4
Figure 4
Thermal denaturation of (A) S11a, (B) S11b, and (C) S11c. Comparison of the fluorescence melting profiles of S11a, S11b and S11c with the 10mer and 20mer peptides.
Figure 5
Figure 5
(A–C) Percentage of positive events of MUC1/Y in SK-OV-3, A498, HT-29, DU145, A549, PC-3, Calu-6, MCF-7, MDA-MB-231 and OVCAR-3 using the aptamers (A) S11a, (B) S11b, and (C) S11c. (D) Representative immunofluorescence of MCF-7, DU145, Calu-6, and A498 cells stained with control-Cy3 (aptamer SP68; selected against a different/non-MUC1 tumour marker peptide), S11a-Cy3 or S11b-Cy3, * p < 0.05.
Figure 6
Figure 6
(A) Radiochemical purity of S11b-99mTc aptamer at 0, 2, 4 and 6 h post-labelling in saline. (B) Tumour growth curve of MDA-MB-231 tumour-bearing NOD-scid mice control (red) and treated with 100 mg/Kg of S11b aptamer (green). Data are the mean (S.D.), n = 5. Data were analysed by a non-linear regression model. **** p < 0.05.
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