Lipoproteins LDL versus HDL as nanocarriers to target either cancer cells or macrophages
Tarik Hadi, Christophe Ramseyer, Thomas Gautier, Pierre-Simon Bellaye, Tatiana Lopez, Antonin Schmitt, Sarah Foley, Semen Yesylevskyy, Thibault Minervini, Romain Douhard, Lucile Dondaine, Lil Proukhnitzky, Samir Messaoudi, Maeva Wendremaire, Mathieu Moreau, Fabrice Neiers, Bertrand Collin, Franck Denat, Laurent Lagrost, Carmen Garrido, Frederic Lirussi
Tarik Hadi, Christophe Ramseyer, Thomas Gautier, Pierre-Simon Bellaye, Tatiana Lopez, Antonin Schmitt, Sarah Foley, Semen Yesylevskyy, Thibault Minervini, Romain Douhard, Lucile Dondaine, Lil Proukhnitzky, Samir Messaoudi, Maeva Wendremaire, Mathieu Moreau, Fabrice Neiers, Bertrand Collin, Franck Denat, Laurent Lagrost, Carmen Garrido, Frederic Lirussi
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Research Article Oncology Therapeutics

Lipoproteins LDL versus HDL as nanocarriers to target either cancer cells or macrophages

Abstract

In this work, we have explored natural unmodified low- and high-density lipoproteins (LDL and HDL, respectively) as selective delivery vectors in colorectal cancer therapy. We show in vitro in cultured cells and in vivo (NanoSPECT/CT) in the CT-26 mice colorectal cancer model that LDLs are mainly taken up by cancer cells, while HDLs are preferentially taken up by macrophages. We loaded LDLs with cisplatin and HDLs with the heat shock protein-70 inhibitor AC1LINNC, turning them into a pair of “Trojan horses” delivering drugs selectively to their target cells as demonstrated in vitro in human colorectal cancer cells and macrophages, and in vivo. Coupling of the drugs to lipoproteins and stability was assessed by mass spectometry and raman spectrometry analysis. Cisplatin vectorized in LDLs led to better tumor growth suppression with strongly reduced adverse effects such as renal or liver toxicity. AC1LINNC vectorized into HDLs induced a strong oxidative burst in macrophages and innate anticancer immune response. Cumulative antitumor effect was observed for both drug-loaded lipoproteins. Altogether, our data show that lipoproteins from patient blood can be used as natural nanocarriers allowing cell-specific targeting, paving the way toward more efficient, safer, and personalized use of chemotherapeutic and immunotherapeutic drugs in cancer.

Authors

Tarik Hadi, Christophe Ramseyer, Thomas Gautier, Pierre-Simon Bellaye, Tatiana Lopez, Antonin Schmitt, Sarah Foley, Semen Yesylevskyy, Thibault Minervini, Romain Douhard, Lucile Dondaine, Lil Proukhnitzky, Samir Messaoudi, Maeva Wendremaire, Mathieu Moreau, Fabrice Neiers, Bertrand Collin, Franck Denat, Laurent Lagrost, Carmen Garrido, Frederic Lirussi

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Figure 5

In vivo effects of cisplatin vectorization in LDL.

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In vivo effects of cisplatin vectorization in LDL. (A) LDL and HDL were ...
(A) LDL and HDL were purified by density gradient ultracentrifugation. Total cholesterol in lipoproteins was adjusted to 1 mM. Lipoproteins were then incubated with cisplatin (10 mg/mL) for 3 hours at 37°C and then submitted to dialysis. Total cisplatin concentration in lipoproteins was then measured by GF-AAS. (B) For cisplatin exchanges, after integration, cisplatin-bound LDL or cisplatin-bound HDL (0.5 μM) were incubated for 24 hours at 37°C with native HDL or LDL (0.5 μM), respectively. LDL and HDL were purified by density gradient ultracentrifugation, and total cisplatin concentration in lipoproteins was then measured. Data are represented as mean value ± SEM. n = 4, ***P < 0.001, ***P < 0.0001. (C) For the antitumor effect, HCT116 cells were treated for 48 hours with cisplatin alone or vectorized in LDL (25 μM final cisplatin concentration). Cell number is represented as mean percentage value versus nontreated ± SEM. n = 4, ***P < 0.001 versus nontreated. (D) For macrophages activation, human M2 macrophages were treated for 2 hours with cisplatin alone or vectorized in LDL (25 μM final cisplatin concentration). Percentages of ROS+ macrophages are represented as mean values ± SEM. n = 4, **P < 0.01. (E) Balb/c mice were injected with CT-26 colorectal cancer cells (1 × 106 cells/mice, s.c.). At the indicated times, mice were treated either with LDL-PBS (100 μM cholesterol), cisplatin (1.5 mg/kg), or LDL-Cis (100 μM cholesterol, 1.5 mg/kg cisplatin). Five mice/group (n = 3). (F) Tumor volume was measured every 3 days and represented as mean value ± SEM. *P < 0.05, **P < 0.01; arrow indicates first injection. (G) Apoptosis and macrophage infiltration were determined in histological slides labeled with a cleaved caspase-3 antibody (green) and a F4/80 antibody (red), and with DAPI. Images were chosen in random fields and are representative of 5 images taken for each condition. n = 5. Scale bar: 50 μm. (H) ROS production in tumors was measured in histological slides by DAPI/DHE staining. Images, taken in random fields, are representative of 5 different ones taken for each condition. n = 5. Scale bar: 50 μm. (I–K) Quantifications of the immunofluorescence intensity of cleaved caspase-3 (I), F4/80 (J), and DHE (K). Data are represented as mean increase versus PBS ± SEM. n = 4. *P < 0.05, **P < 0.01, ****P < 0.0001. P values were calculated using 1-way ANOVA (A, F, and I–K) or 2-tailed unpaired t tests (C and D).