Dendrimers as Drug Nanocarriers: The Future of Gene Therapy and Targeted Therapies in Cancer

Synthetic polymers, such as dendrimers, play a critical role in pharmaceutical discovery and development. Advances in the application of nanotechnology in medicine have given rise to multifunctional “smart” nanocarriers that can deliver one or more therapeutic agents safely and selectively to cancer cells, including intracellular gene-specific targeting. Dendrimers with their 3D nanopolymeric architectures are highly attractive class of drug and gene delivery vector. Advances in understanding and manipulating genes gave scientists a tool to make changes in people DNA to prevent or treat diseases. Over the past decade, gene therapy has been in use in clinical trials. The inactivation of the tumor suppressor genes is the main idea of the development of gene therapy in the cancer treatment. Broad spectrum of delivery concepts, including viral vectors, liposomes, cationic polymers and dendrimers, cell-penetrating peptides and gold and magnetic nanoparticles have been investigated. A well-designed vector is the most desirable approach to increase the safety of gene therapy, which is still in its infancy stages in cancer research. More experimental and clinical trials are focused on well-designed and effective doses of vectors that are essential for therapeutic efficacy of gene therapy for its potential in clinical use against a wide variety of cancers

Affecting NF-κB cell signaling pathway in chronic lymphocytic leukemia by dendrimers-based nanoparticles

Abstract The complex genetic diversity of chronic lymphocytic leukemia (CLL) makes it difficult to determine the effective and durable therapy beneficial to patients. During the several past years' significant insights in the biology of the disease and its treatment have been made, allowing for the identification of promising novel therapeutic agents. The investigation of signaling pathways to understand the biological character of CLL together with the development of molecular profiling is key in personalized approach in therapy for this disease. As it was already proven, maltotriose (M3) modified fourth generation poly(propylene imine) dendrimers(PPI-G4) modulate BCR, TRAIL and WNT signaling pathway gene expression in CLL cells and strongly influence their survival by inducing apoptosis and inhibiting proliferation. The aim of this study was to evaluate the influence of PPI-G4-M3 dendrimers on NFκB pathway gene expression in CLL (MEC-1) cells with 60 K microarray, as it is one of the major factors in the pathogenesis of B-cell neoplasms. The findings were compared with those obtained with Fludarabine (FA) and the results indicate that PPI-G4-M3 dendrimers affect the expression of the examined genes and exert comparable effect on the CLL cells to FA. Dendrimers are one of the most potent groups of nanometer-sized macromoleculesfor closing the gap between the present ineffective treatment and the future effective personalized therapy due to their potential versatile biological propertie

POTENTIAL GENETIC AGENT BFL1 FOR TARGETED THERAPY IN CHRONIC LYMPHOCYTIC LEUKEMIA

Background: Many prognostic factors have been identified in chronic lymphocytic leukemia (CLL) but new ones are still desired. The biological characterization of CLL is now being translated into novel treatment strategies. One new prognostic factor, and therapeutic target, may be BFL1. It is both a serum and a molecular marker that contributes to the progression of CLL and its resistance to chemotherapy. The aim of this study was to evaluate the prognostic value of BFL1 and to assess its correlation with other known prognostic markers in CLL for the cladribine and cyclophosphamide regimen (CC). Methods: qPCR TaqMan® Low Density Array was used for gene expression measurements. Assessment of CD38, ZAP70 and BFL-1 proteins expression was done by means of flow cytometry. Serum TK activity was measured by immunoassay. Results: Protein BFL1 expression was found to be significantly higher in CLL patients than healthy volunteers (p=0.001). Moreover its level was significantly higher in patients with no response (NR) to CC therapy (p=0.009). The expression of BFL1 was considerably down regulated during CC treatment and BFL1 mRNA levels were inversely correlated with apoptotic response. In addition, protein BFL1 expression was found to be similar to thymidine kinase (TK) concentration regarding treatment response. As far as other markers are concerned, a positive correlation was identified between BFL1 and TK (r=0.52, p=0.01). Conclusions: Our findings suggest that BFL1 contributes to chemoresistance and may be a co-existing prognostic factor in CLL in the future

Dendritic glycopolymers based on dendritic polyamine scaffolds: view on their synthetic approaches, characteristics and potential for biomedical applications

In this review we highlight the potential for biomedical applications of dendritic glycopolymers based on polyamine scaffolds. The complex interplay of the molecular characteristics of the dendritic architectures and their specific interactions with various (bio)molecules are elucidated with various examples. A special role of the individual sugar units attached to the dendritic scaffolds and their density is identified, which govern ionic and H-bond interactions, and biological targeting, but to a large extent are also responsible for the significantly reduced toxicity of the dendritic glycopolymers compared to their polyamine scaffolds. Thus, the application of dendritic glycopolymers in drug delivery systems for gene transfection but also as therapeutics in neurodegenerative diseases has great promisePublikacja w ramach programu Royal Society of Chemistry "Gold for Gold" 2014 finansowanego przez Uniwersytet Łódzk

Electrophoretic characteristics of metalloproteinase’s activity of serum of patients with chronic lymphocytic leukemia – preliminary data

Metalloproteinases play an important role in the development and metastasis of many cancers. Their activity is also an important component of tumorgenesis associated processes such as angiogenesis, decreased apoptosis, or unlimited proliferation of pathological cells. In this study we tried to estimate a differences of metalloproteinase activity digesting the gelatin in the sera of patients with chronic lymphocytic leukemia and healthy people, by the zymographic technique. To confirm that the gelatinolytic activity originated from the metalloproteinases their specific inhibitors: phenanthroline and ethylene– diaminetetraacetic acid were used. In patient’s sera a zymographic analysis revealed the presence of additional activity. The first of them are located in a region corresponding to a molecular weight of approximately 240 kDa, probably corresponds to the dimer of proMMP–9. Another two active fractions present in the sera of patients suffering from leukemia corresponded to a molecular weight of about 110 and 130 kDa probably represents a complex of proMMP–9 with lipocain. In a control sera, only one activity could be observed exhibiting a molecular weight of about 110 kDa, which is stronger than corresponding fraction in patient’s sera. The biggest difference between the two investigated sera was gelatynolytic activity located in the region of a molecular weight of about 94 kDa, which probably corresponded to proMMP–9. In some leukemic sera this activity was several times higher compared to the control samples, in which there was a constant and relatively low level of it. Despite significant activity of proMMP–9 in sera of patients with CLL no biologically active equivalent band of molecular weight 84 kDa were detected. The fractions which corresponded to different forms of MMP–2 (72, 64 kDa) were present in the sera of tumor and control, although proMMP–2 was more strongly expressed in the control samples

Electrophoretic method of cysteine proteinase identification in the sera of patients with chronic lymphocytic leukemia (CLL) based on biotinylated iodoacetamide

Wstęp: Proteinazy cysteinowe są enzymami regulującymi liczne procesy fizjologiczne oraz patologiczne w organizmie człowieka. Zaburzenie ich aktywności może przyczyniać się do wystąpienia wielu chorób. Pełnią one ważną rolę w procesie kancerogenezy, uczestnicząc w inwazji, transformacji nowotworowej, angiogenezie, apoptozie oraz powstawaniu przerzutów. Celem pracy było opracowanie elektroforetycznej metody identyfikacji proteinaz cysteinowych w surowicach pacjentów z przewlekłą białaczką w oparciu o jodoacetamid biotynylowany. Materiał i metody: Badania wstępne przeprowadzono na handlowym preparacie papainy (EC 3.4.22.2), dobrze poznanej i szeroko stosowanej roślinnej proteinazy cysteinowej o masie cząsteczkowej 23,4 kDa. Badania wykonano na próbach surowicy uzyskanych z pełnej krwi pobranej od pacjentów chorych na przewlekłą białaczkę limfocytową (PBL) oraz surowicach kontrolnych uzyskanych od dawców. Surowice po wstępnej inkubacji z jodoacetamidem mieszano z buforem do prób i poddawano rozdziałowi elektroforetycznemu w żelu poliakrylamidowym zawierającym dodecylosiarczan sodu (SDS–PAGE). Rozdzielone elektroforetycznie białka po transferze na nitrocelulozową membranę, poddawano dalszej analizie za pomocą streptawidyny skoniugowanej z peroksydazą chrzanową (HRP). Użycie substratu dla HRP, czterochlorowodorku 3,3’–diaminobenzydynyny (DAB), umożliwiało identyfikację biotynylowanego jodoacetamidu, a przez to związanych z nim w sposób nieodwracalny proteinaz cysteinowych obecnych w badanych surowicach. Wyniki: Analiza porównawcza surowic pobranych od pacjentów chorych na przewlekłą białaczkę limfocytową w odniesieniu do surowic kontrolnych pozwoliła na identyfikację dodatkowego białka o charakterze proteinazy cysteinowej o masie cząsteczkowej wynoszącej około 37 kDa, które nie występowało lub było obecne w niewielkiej ilości w surowicach kontrolnych. Wnioski: Opracowana metoda umożliwia wykrywanie proteinaz cysteinowych w surowicach kontrolnych, jak i u pacjentów chorych na przewlekłą białaczkę limfocytową.Introduction: Cysteine proteases are enzymes that regulate numerous physiological and pathological processes in the human body. Disorders of their activity can lead to a number of diseases. They play an important role in the process of carcinogenesis, participating in the invasion, transformation, angiogenesis, apoptosis and metastasis. The aim of this study was to elaborate the electrophoretic method of cysteine proteinases identification in the sera of patients suffering from chronic lymphocytic leukemia based on biotinylated iodoacetamide. Material and methods: Preliminary studies were carried out on the commercially available papain (EC 3.4.22.2) well known and widely used plant cysteine protease with a molecular weight 23,4 kDa. The study was conducted on the blood samples taken from patients with chronic lymphocytic leukemia (CLL) and control sera from healthy donors. The sera after the preincubation with iodoacetamide were mixed with the sample buffer followed by electrophoresis on polyacrylamide gel containing sodium dodecyl sulfate (SDS–PAGE). The separated proteins were electrophoretically transferred to the nitrocellulose membranes and subjected to the further analysis using streptavidin conjugated with horseradish peroxidase (HRP). The use of substrate for HRP 3,3’- diaminobenzidine tetrachloride (DAB) allows the biotinylated iodoacetamide and thereby cysteine proteinase identification. Results: The comparative analysis of the sera from the patients with chronic lymphocytic leukemia and the control sera led to the identification of additional protein with a cysteine protease characteristic having a molecular weight of about 37 kDa, which did not occur or was present in a smaller amount of the control sera. Conclusions: The developed method allows the detection of cysteine proteases which are present in the control sera and the sera of patients with chronic lymphocytic leukemia