ABSTRACT
Gene silencing through RNA interference (RNAi) is a promising therapeutic approach for a wide range of disorders, including cancer. Non-viral gene therapy, using specific siRNAs against BCR-ABL1, can be a supportive or alternative measure to traditional chronic myeloid leukemia (CML) tyrosine kinase inhibitor (TKIs) therapies, given the prevalence of clinical TKI resistance. The main challenge for such approaches remains the development of the effective delivery system for siRNA tailored to the specific disease model. The purpose of this study was to examine and compare the efficiency of endosomolytic cell penetrating peptide (CPP) EB1 and PEG2000-decorated cationic liposomes composed of polycationic lipid 1,26-bis(cholest-5-en-3-yloxycarbonylamino)-7,11,16,20-tetraazahexacosane tetrahydrochloride (2Ð¥3) and helper lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) for anti-bcr-abl siRNA delivery into the K562 human CML cell line. We show that both EB1 and 2Ð¥3-DOPE-DSPE-PEG2000 (0.62 % mol.) liposomes effectively deliver siRNA into K562 cells by endocytic mechanisms, and the use of liposomes leads to more effective inhibition of expression of the targeted gene (BCR-ABL1) and cancer cell proliferation. Taken together, these findings suggest that PEG-decorated cationic liposomes mediated siRNA delivery allows an effective antisense suppression of certain oncogenes, and represents a promising new class of therapies for CML.
Subject(s)
Cell-Penetrating Peptides , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Liposomes , RNA, Small Interfering , Humans , Liposomes/chemistry , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/therapy , Cell-Penetrating Peptides/chemistry , Cell-Penetrating Peptides/pharmacology , RNA, Small Interfering/genetics , RNA, Small Interfering/administration & dosage , Fusion Proteins, bcr-abl/genetics , Fusion Proteins, bcr-abl/metabolism , Fusion Proteins, bcr-abl/antagonists & inhibitors , Cell Proliferation/drug effects , Polyethylene Glycols/chemistry , K562 Cells , Phosphatidylethanolamines/chemistry , Cations/chemistryABSTRACT
PURPOSE: Metronomic chemotherapy (MC) is a promising approach where, in contrast to the conventional maximal tolerated dose (MTD) strategy, regular fractionated doses of the drug are used. This approach has proven its efficacy, although drug dosing and scheduling are often chosen empirically. Pharmacokinetic/pharmacodynamic (PK/PD) models provide a way to choose optimal protocols with computational methods. Existing models are usually too complicated and are valid for only a subset of drug schedules. To address this issue, we propose herein a simple model that can describe MC and MTD regimens simultaneously. METHODS: The minimal model comprises tumor suppression due to antiangiogenic drug effect together with a cell-kill term, responsible for its cytotoxicity. The model was tested on data obtained on tumor-bearing mice treated with gemcitabine in ether MTD, MC, or combined (MTD + MC) regimens. RESULTS: We conducted a number of tests in which data were divided in various ways into training and validation sets. The model successfully described different trends in the MTD and MC regimens. With parameters obtained by fitting the model to MTD data, the simulations correctly predicted trends in both the MC and combined therapy groups. CONCLUSION: Our results demonstrate that the proposed model presents a minimal yet efficient tool for modeling outcomes in different treatment regimens in mice. We hope that this model has the potential for use in clinical practice in the development of patient-specific chemotherapy scheduling protocols based on observed treatment response.
Subject(s)
Antineoplastic Agents/administration & dosage , Antineoplastic Agents/pharmacokinetics , Carcinoma, Ehrlich Tumor/drug therapy , Administration, Metronomic , Animals , Carcinoma, Ehrlich Tumor/pathology , Deoxycytidine/administration & dosage , Deoxycytidine/analogs & derivatives , Deoxycytidine/pharmacokinetics , Female , Maximum Tolerated Dose , Mice , Models, Theoretical , Reproducibility of Results , GemcitabineABSTRACT
Prorocentrum minimum is a potentially toxic marine dinoflagellate that often forms massive blooms in estuarine and coastal sea waters. In this study, the life cycle of P. minimum was investigated and sexual reproduction in culture was described for the first time. Morphology of the mitotic stages was revised and several distinguishing features from sexual steps were described. The sexual reproductive stages were observed in the stationary culture and compared with a well-studied closely related species, Prorocentrum micans. Prorocentrum minimum has a haplontic life cycle and homothallic sexual process. The gametes were isogamous and morphologically indistinguishable from the vegetative cells. Unlike P. micans, P. minimum isogametes fused, but did not conjugate, partially reorganizing their cell coverings. Newly formed planozygotes were distinguished by their irregular shape and a large asymmetrically located nucleus. No long-term resting cyst stages (hypnozygotes) were documented. The late planozygotes underwent meiosis and formed tetrads of cells. The second meiotic division could be delayed or arrested in one of the daughter nuclei leading to formation of trinucleate cells with three pairs of flagella. So, similar to P. micans, P. minimum may have two possible scenarios of sexual division: (a) formation of a four-cell stage through two successive divisions or (b) asynchronous divisions of the zygote. Changes in the DNA content were confirmed by quantitative image cytometry.
