Preclinical progress and first translational steps for a liposomal chemotherapy protocol against adrenocortical carcinoma

Abstract

Systemic therapy of adrenocortical carcinoma (ACC) is limited by heterogeneous tumor response and adverse effects. Recently, we demonstrated anti-tumor activity of LEDP-M (etoposide, liposomal doxorubicin, liposomal cisplatin, mitotane), a liposomal variant of EDP-M (etoposide, doxorubicin, cisplatin, mitotane). To improve the therapeutic efficacy and off-target profiles of the clinical gold standard EDP-M, we investigated liposomal EDP-M regimens in different preclinical settings and in a small number of ACC patients with very advanced disease. Short- and long-term experiments were performed on two ACC models (SW-13 and SJ-ACC3) in vivo We evaluated the anti-tumoral effects and off-target profiles of EDP-M, LEDP-M and a novel regimen L(l)EDP-M including liposomal etoposide. Furthermore, the role of plasma microRNA-210 as a therapeutic biomarker and first clinical data were assessed. Classical and liposomal protocols revealed anti-proliferative efficacy against SW-13 (EDP-M P < 0.01; LEDP-M: P < 0.001; L(l)EDP-M: P < 0.001 vs controls), whereas in SJ-ACC3, only EDP-M (P < 0.05 vs controls) was slightly effective. Long-term experiments in SW-13 demonstrated anti-tumor efficacy for all treatment schemes (EDP-M: P < 0.01, LEDP-M: P < 0.05, L(l)EDP-M P < 0.001 vs controls). The analysis of pre-defined criteria leading to study termination revealed significant differences for control (P < 0.0001) and EDP-M (P = 0.003) compared to L(l)EDP-M treatment. Raising its potential for therapy monitoring, we detected elevated levels of circulating microRNA-210 in SW-13 after LEDP-M treatment (P < 0.05). In contrast, no comparable effects were detectable for SJ-ACC3. However, overall histological evaluation demonstrated improved off-target profiles following liposomal regimens. The first clinical data indicate improved tolerability of liposomal EDP-M, thus confirming our results. In summary, liposomal EDP-M regimens represent promising treatment options to improve clinical treatment of ACC.

Targeting the multidrug transporter Patched potentiates chemotherapy efficiency on adrenocortical carcinoma in vitro and in vivo

“…All experiments were carried out following protocols approved by the Regierung von Oberbayern and in accordance with the German guidelines for animal studies. H295R xenografts were induced as described before . For short‐term experiments, tumor‐bearing mice ( n = 4–5) were treated with one therapeutic cycle which consisted of either liposomal doxorubicin (lipdxr) (6 mg/kg body weight, intravenously on Days 4 and 10), P375 (2 mg/kg body weight, intraperitoneally on Days 1–11) or a combination of lipdxr and P375 (P375: 2 mg/kg body weight intraperitoneally on Days 1–11 plus lipdxr 6 mg/kg body weight intravenously on Days 4 and 10).…”

“…Afterwards, tumors were excised and subsequently paraffin embedded. The slide preparation of these paraffin embedded tissues as well as the quantification of the number of apoptotic tumor cells, using the colorimetric DeadEND TUNEL System (Promega, Mannheim, Germany), were performed as described elsewhere . For long‐term therapeutic protocols, mice ( n = 7–8) received two treatment cycles with a therapy‐free interval of 6 days in between.…”

A mikro-RNS-ek patogenetikai és diagnosztikai szerepe mellékvesekéreg-carcinomában

“…Munkacsoportunk két egér-xenograftmodellben vizsgálta a keringő mikro-RNS-ek és a daganatok kezelésének kapcsolatát. A már sokszor említett keringő miR-483-5p csökkenését észleltük 9-cisz-retinsav-kezelésre egy NCI-H295R mellékvesekéregcarcinoma-sejtvonalból előállí-tott xenograftmodellen [49], míg egy SW-13-as mellék-vesekéregcarcinoma-sejtvonalon alapuló modellben a szöveti hypoxiát jelző miR-210 növekedését észleltük liposzomális etopozid-ciszplatin-doxorubicin-mitotán kezelés hatására [50].…”

Update on microRNA as Biomarkers of Adrenocortical Cancer: Perspective on Circulating microRNA

“…In fact, we have found much fewer significantly differentially expressed microRNAs in extracellular vesicle preparations than in unfractionated plasma that might also confirm our hypothesis that extracellular vesicles could be more specific as a source of microRNAs [Perge et A further potential clinical application of circulating microRNAs could be disease-progression monitoring. We have studied circulating microRNA markers in two ACC xenograft models: in an NCI-H295R xenograft model, we have noted that treatment with 9-cis retinoic acid significantly decreased circulating hsa-miR-483-5p expression [18]; in contrast, circulating hsa-miR-483-5p levels have not been affected by shortterm cytotoxic liposomal-EDP-M (etoposidedoxorubicin-cisplatin-mitotane) chemotherapy in either SW-13 or SJ-ACC3 (a pediatric ACC model) xenografts [19]. Another microRNA, hsamiR-210 that is a major hypoxamiR [20] and was found to be overexpressed as tissue microRNA in ACC, as well [7], turned out to display elevated plasma levels after liposomal EDP-M chemotherapy in the SW-13 model [19].…”

“…We have studied circulating microRNA markers in two ACC xenograft models: in an NCI-H295R xenograft model, we have noted that treatment with 9-cis retinoic acid significantly decreased circulating hsa-miR-483-5p expression [18]; in contrast, circulating hsa-miR-483-5p levels have not been affected by shortterm cytotoxic liposomal-EDP-M (etoposidedoxorubicin-cisplatin-mitotane) chemotherapy in either SW-13 or SJ-ACC3 (a pediatric ACC model) xenografts [19]. Another microRNA, hsamiR-210 that is a major hypoxamiR [20] and was found to be overexpressed as tissue microRNA in ACC, as well [7], turned out to display elevated plasma levels after liposomal EDP-M chemotherapy in the SW-13 model [19]. Considering the lack of suitable blood-borne markers of ACC progression at present, if validated in the human "Given the major difficulties in the diagnosis of ACC including the lack of an applicable preoperative blood marker of malignancy, and difficulties of imaging and histological analysis, circulating microRNAs might be envisaged as potential, novel minimally invasive markers of malignancy.…”