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. 2022 May;9(13):e2105506.
doi: 10.1002/advs.202105506. Epub 2022 Mar 4.

Integrated Design of a Membrane-Lytic Peptide-Based Intravenous Nanotherapeutic Suppresses Triple-Negative Breast Cancer

Charles H Chen  1   2 , Yu-Han Liu  3 , Arvin Eskandari  1 , Jenisha Ghimire  4 , Leon Chien-Wei Lin  3 , Zih-Syun Fang  3 , William C Wimley  4 , Jakob P Ulmschneider  5 , Kogularamanan Suntharalingam  6 , Che-Ming Jack Hu  3 , Martin B Ulmschneider  1
Affiliations

Integrated Design of a Membrane-Lytic Peptide-Based Intravenous Nanotherapeutic Suppresses Triple-Negative Breast Cancer

Charles H Chen et al. Adv Sci (Weinh). 2022 May.

Abstract

Membrane-lytic peptides offer broad synthetic flexibilities and design potential to the arsenal of anticancer therapeutics, which can be limited by cytotoxicity to noncancerous cells and induction of drug resistance via stress-induced mutagenesis. Despite continued research efforts on membrane-perforating peptides for antimicrobial applications, success in anticancer peptide therapeutics remains elusive given the muted distinction between cancerous and normal cell membranes and the challenge of peptide degradation and neutralization upon intravenous delivery. Using triple-negative breast cancer as a model, the authors report the development of a new class of anticancer peptides. Through function-conserving mutations, the authors achieved cancer cell selective membrane perforation, with leads exhibiting a 200-fold selectivity over non-cancerogenic cells and superior cytotoxicity over doxorubicin against breast cancer tumorspheres. Upon continuous exposure to the anticancer peptides at growth-arresting concentrations, cancer cells do not exhibit resistance phenotype, frequently observed under chemotherapeutic treatment. The authors further demonstrate efficient encapsulation of the anticancer peptides in 20 nm polymeric nanocarriers, which possess high tolerability and lead to effective tumor growth inhibition in a mouse model of MDA-MB-231 triple-negative breast cancer. This work demonstrates a multidisciplinary approach for enabling translationally relevant membrane-lytic peptides in oncology, opening up a vast chemical repertoire to the arms race against cancer.

Keywords: anticancer peptides; drug resistance; membrane-active anticancer agents; multicellular tumor spheroids; murine models; nanoparticles; triple negative breast cancer.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Schematic diagram of integrated design of peptide‐based nanomedicine in cancer treatment. The process of nanomedicine development involves peptide drug design, evaluation against both 2D and 3D cell cultures, and nanomedicine formulation for animal study.
Figure 2
Figure 2
Compounds selectivity for cancer cells in vitro. a) Comparison of compound IC50 against cancerous (HMLER) and noncancerous (MCF‐10A) human breast epithelial cells in a 2D in vitro culture model. b) Comparison of IC50 epithelial breast cancer stem cell (HMLER‐shEcad) and noncancerous MCF‐10A. c) Comparison of IC50 values for doxorubicin, salinomycin, and EEK against HMLER, HMLER‐shEcad, MCF‐10A, and HEK293T. d) IC50 comparison of HMLER and HEK293T. e) IC50 comparison of HMLER‐shEcad and HEK293T. f) Comparison of selectivity for HMLER and HMLER‐shEcad.
Figure 3
Figure 3
Comparison of compound activity against cancer stem cell spheroids and evaluation of resistance formation. a) Dose‐dependent cell viability of HMLER‐shEcad (cancer stem cell) tumorspheres treated with either doxorubicin, DHK, EEK, or the two ACP D‐enantiomers d‐DHK and d‐EEK. b) Dose‐dependent cell viability of MCF‐10A (non‐tumorigenic cell) mammospheres treated with the same compounds. c) IC50 (dark color) and IC90 (light color) of all compounds tested against HMLER‐shEcad tumorspheres (blue) and MCF‐10A mammospheres (red). d) IC50 (dark color) and IC90 (light color) of doxorubicin, DHK, and d‐DHK, in 2D HMLER‐shEcad cells after prolonged exposure to vehicle (blue) and doxorubicin or d‐DHK (green). e) Dose‐dependent cell viability of 2D HMLER‐shEcad after 28 days of co‐incubation with 0.4 µm d‐DHK or vehicle. The scale bar is 100 µm. f) Dose‐dependent cell viability of 2D HMLER‐shEcad cells after 10 days of co‐incubation with either 30 nm doxorubicin or vehicle.
Figure 4
Figure 4
ACP membrane pore structures and membrane perforation mechanism. Molecular dynamics simulations reveal the full atomic details of a) spontaneous ACP membrane adsorption, b) insertion, and c) pore formation (shown is a large, heterogeneous, fully water‐filled EEK pore). d,e) Bound peptides form an ensemble of transient pores of 2–16 peptides (top) that conduct both water (middle) and ions (bottom) across the membrane.
Figure 5
Figure 5
Development of EEK nanoparticles. a) Schematic illustration of L‐EEK NPs and NPs preparation. b) Transmission electron microscopy images of L‐EEK NPs. Scale bars are 200 nm (black) and 20 nm in the inset (white). c) Dynamic light scattering characterizations of L‐EEK NPs (n = 3) and control NPs without L‐EEK cargo. d) The comparative haemolytic activities of free L‐form EEK, L‐EEK NPs, and control NPs. e) Assessment of cell viability by CCK‐8 assay with L‐EEK, L‐EEK NPs, and control NPs treatment against breast cancer cell lines (MCF‐7, MDA‐MB‐231, MDA‐MB‐453, and ZR‐75‐1). IC50 values of L‐EEK and L‐EEK NPs against breast cancer cells are in green and red, respectively. f) Schematics of the mouse model of MDA‐MB‐231 triple‐negative breast cancer with control nanoparticles and EEK peptide nanoparticles treatment schedule. g) Efficient inhibition of cancer growth with L‐form EEK NPs treatment. Upon establishment of palpable tumors on day 11 following subcutaneous inoculation with MDA‐MB‐231 (4 × 106 cells), mice were treated with 10 mg kg−1 per dose of EEK‐NPs or equivalent doses of control NPs over a 14‐day treatment period. Tumor volumes were monitored. ***p < 0.005 (n = 4). h) Images of MDA‐MB‐231 tumors on day 33 after the onset of L‐EEK NPs and control NPs treatments. i) Kaplan–Meier curve of mice survival following tumor inoculation over an observation period of 120 days. Mouse survival is defined as tumor size below 1000 mm3 (n = 4, *p < 0.05).
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References

    1. a) Marx J., Science 2007, 317, 1029; - PubMed
    2. b) Nguyen L. V., Vanner R., Dirks P., Eaves C. J., Nat. Rev. Cancer 2012, 12, 133; - PubMed
    3. c) Pattabiraman D. R., Weinberg R. A., Nat. Rev. Drug Discovery 2014, 13, 497; - PMC - PubMed
    4. d) Plaks V., Kong N., Werb Z., Cell Stem Cell 2015, 16, 225; - PMC - PubMed
    5. e) Kaiser J., Science 2015, 347, 226. - PubMed
    1. Dean M., Fojo T., Bates S., Nat. Rev. Cancer 2005, 5, 275. - PubMed
    1. Guha S., Ghimire J., Wu E., Wimley W. C., Chem. Rev. 2019, 119, 6040. - PMC - PubMed
    1. Lazzaro B. P., Zasloff M., Rolff J., Science 2020, 368, eaau5480. - PMC - PubMed
    1. Chen C. H., Lu T. K., Antibiotics 2020, 9, 24.

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