Intra- and Inter-Species Variability in Urinary N7-(1-Hydroxy-3-buten-2-yl)guanine Adducts Following Inhalation Exposure to 1,3-Butadiene.

1,3-Butadiene is a known carcinogen primarily targeting lymphoid tissues, lung, and liver. Cytochrome P450 activates butadiene to epoxides which form covalent DNA adducts that are thought to be a key mechanistic event in cancer. Previous studies suggested that inter-species, -tissue, and -individual susceptibility to adverse health effects of butadiene exposure may be due to differences in metabolism and other mechanisms. In this study, we aimed to examine the extent of inter-individual and inter-species variability in the urinary N7-(1-hydroxy-3-buten-2-yl)guanine (EB-GII) DNA adduct, a well-known biomarker of exposure to butadiene. For a population variability study in mice, we used the collaborative cross model. Female and male mice from five strains were exposed to filtered air or butadiene (590 ppm, 6 h/day, 5 days/week for 2 weeks) by inhalation. Urine samples were collected, and the metabolic activation of butadiene by DNA-reactive species was quantified as urinary EB-GII adducts. We quantified the degree of EB-GII variation across mouse strains and sexes; then, we compared this variation with the data from rats (exposed to 62.5 or 200 ppm butadiene) and humans (0.004-2.2 ppm butadiene). We show that sex and strain are significant contributors to the variability in urinary EB-GII levels in mice. In addition, we find that the degree of variability in urinary EB-GII in collaborative cross mice, when expressed as an uncertainty factor for the inter-individual variability (UFH), is relatively modest (≤threefold) possibly due to metabolic saturation. By contrast, the variability in urinary EB-GII (adjusted for exposure) observed in humans, while larger than the default value of 10-fold, is largely consistent with UFH estimates for other chemicals based on human data for non-cancer endpoints. Overall, these data demonstrate that urinary EB-GII levels, particularly from human studies, may be useful for quantitative characterization of human variability in cancer risks to butadiene.

In vivo localization and postmortem stability of benzo[a]pyrene-DNA adducts.

DNA adducts of carcinogenic polycyclic aromatic hydrocarbons (PAHs) play a critical role in the etiology of gastrointestinal tract cancers in humans and other species orally exposed to PAHs. Yet, the precise localization of PAH-DNA adducts in the gastrointestinal tract, and the long-term postmortem PAH-DNA adduct stability are unknown. To address these issues, the following experiment was performed. Mice were injected intraperitoneally with the PAH carcinogen benzo[a]pyrene (BP) and euthanized at 24 h. Tissues were harvested either at euthanasia (0 time), or after 4, 8, 12, 24, 48, and 168 hr (7 days) of storage at 4°C. Portions of mouse tissues were formalin-fixed, paraffin-embedded, and immunohistochemically (IHC) evaluated by incubation with r7,t8-dihydroxy-t-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE)-DNA antiserum and H-scoring. The remaining tissues were frozen, and DNA was extracted and assayed for the r7,t8,t9-trihydroxy-c-10-(N 2 -deoxyguanosyl)-7,8,9,10-tetrahydrobenzo[a]pyrene (BPdG) adduct using two quantitative assays, the BPDE-DNA chemiluminescence immunoassay (CIA), and high-performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ES-MS/MS). By IHC, which required intact nuclei, BPdG adducts were visualized in forestomach basal cells, which included gastric stem cells, for up to 7 days. In proximal small intestine villus epithelium BPdG adducts were visualized for up to 12 hr. By BPDE-DNA CIA and HPLC-ES-MS/MS, both of which used DNA for analysis and correlated well (P= 0.0001), BPdG adducts were unchanged in small intestine, forestomach, and lung stored at 4°C for up to 7 days postmortem. In addition to localization of BPdG adducts, this study reveals the feasibility of examining PAH-DNA adduct formation in wildlife species living in colder climates. Environ. Mol. Mutagen. 61:216-223, 2020. © 2019 Wiley Periodicals, Inc.

Targeted delivery of doxorubicin to cancer cells by a cruciform DNA nanostructure composed of AS1411 and FOXM1 aptamers.

OBJECTIVES: Here, a novel cruciform DNA nanostructure was developed for targeted delivery of doxorubicin (Dox), as an anticancer agent, to lung (A549 cells) and breast (4T1 cells) cancer cells. The cruciform DNA nanostructure consisted of AS1411 aptamer as targeting agent and Forkhead Box Protein M1(FOXM1) aptamer as therapeutic agent. METHODS: MTT assay, fluorescence imaging, flow cytometry analysis, and in vivoantitumor efficacy were performed to evaluate the function of the Dox-DNA nanostructure complex. RESULTS: The presented delivery system benefited from tumor targeting, high stability in serum and simple construction. The Dox-DNA nanostructure complex showed a noticeable higher internalization degree into A549 and 4T1 cells (target), overexpressing nucleolin on their cell membranes, compared to CHO cells (nontarget, nucleolin negative). Moreover, the results of MTT assay exhibited that Dox-DNA nanostructure complex significantly decreased cell viability in A549 and 4T1 cells compared to CHO cells, which significantly preserved their viability. Besides, Dox-DNA nanostructure complex significantly reduced tumor growth in tumor-bearing mice in comparison with Dox and DNA nanostructure treatments. CONCLUSION: These findings confirmed that synergistic combination of FOXM1 aptamer and Dox into Dox-DNA nanostructure complex enhanced antitumor effectiveness and reduced toxicity toward nontarget cells, opening up new insights in cancer treatment.

Tumor Acidity/NIR Controlled Interaction of Transformable Nanoparticle with Biological Systems for Cancer Therapy.

Precisely controlling the interaction of nanoparticles with biological systems (nanobio interactions) from the injection site to biological targets shows great potential for biomedical applications. Inspired by the ability of nanoparticles to alter their physicochemical properties according to different stimuli, we explored the tumor acidity and near-infrared (NIR) light activated transformable nanoparticle DATAT-NPIR&DOX. This nanoparticle consists of a tumor acidity-activated TAT [the TAT lysine residues' amines was modified with 2,3-dimethylmaleic anhydride (DA)], a flexible chain polyphosphoester core coencapsulated a NIR dye IR-780, and DOX (doxorubicin). The physicochemical properties of the nanoparticle can be controlled in a stepwise fashion using tumor acidity and NIR light, resulting in adjustable nanobio interactions. The resulting transformable nanoparticle DATAT-NPIR&DOX efficiently avoids the interaction with mononuclear phagocyte system (MPS) ("stealth" state) due to the masking of the TAT peptide during blood circulation. Once it has accumulated in the tumor tissues, DATAT-NPIR&DOX is reactivated by tumor acidity and transformed into the "recognize" state in order to promote interaction with tumor cells and enhance cellular internalization. Then, this nanoparticle is transformed into "attack" state under NIR irradiation, achieving the supersensitive DOX release from the flexible chain polyphosphoester core in order to increase the DOX-DNA interaction. This concept provides new avenues for the creation of transformable drug delivery systems that have the ability to control nanobio interactions.

Interaction of Different Divalent Metal Ions with Lipid Bilayer: Impact on the Encapsulation of Doxorubicin by Lipid Bilayer and Lipoplex Mediated Deintercalation.

In this article, we investigate the influence of different metal ions (Ca2+, Mg2+, and Zn2+) on binding of an anticancer drug doxorubicin (DOX) to DMPC bilayer and lipoplex mediated deintercalation of DOX from DOX-DNA complex. Our study reveals that lipid bilayer in the presence of different metal ions displays much higher binding affinity toward DOX than bare lipid bilayer does. Further, this affinity for a particular metal ion increases linearly with metal ion concentration. The steady state and time-resolved fluorescence studies reveal that binding of DOX with lipid bilayer in the presence of different metal ions varies in the order of Ca2+> Mg2+> Zn2+. The rotational relaxation of DOX in the presence of different metal ions takes place in the same order. We explain these phenomena in the light of alteration of the physical properties brought about by metal ions. Moreover, we find that binding pattern of metal ions with lipid head groups influences the intake of DOX in lipid bilayer. We exploit the binding of DOX with bilayer to study the deintercalation of DOX from DOX-DNA complex. We observe that with increase in metal ion concentration the deintercalation increases. Among all metal ions, Ca2+ appears to be most effective in deintercalation compared to other metal ions. The time-resolved fluorescence anisotropy and circular dichroism measurements indicate that in the presence of Ca2+, lipid bilayer offer strongest interaction with DNA while the same is weakest for Zn2+. This explains the highest percentage of deintercalation of DOX from drug-DNA complex in the presence of Ca2+. Overall the present study demonstrates a new strategy that binding of drug molecules with lipid bilayer and deintercalation of the same from drug-DNA complex can be tuned by modulation of lipid bilayer with different metal ions and their concentration.

RETRACTED: Low density lipoprotein receptor targeted doxorubicin/DNA-Gold Nanorods as a chemo- and thermo-dual therapy for prostate cancer.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the authors and sanctioned by the Editor-in-Chief. The authors found errors in the data presentation - apoptotic statistics and in vivo distribution - which makes the conclusion not representative. The authors express sincere apologies for the error and inconvenience to readers.

Tumor-Penetrating Peptide-Modified DNA Tetrahedron for Targeting Drug Delivery.

DNA self-assembling nanostructure has been considered as a promising candidate as a drug delivery vehicle because of its compactness, mechanical stability, and noncytotoxicity. In this work, we developed functional, multiform DNA nanostructures by appending a tumor-penetrating peptide to tetrahedral DNA nanostructure (p-TDN). This functional structure is able to efficiently increase the rate of uptake of glioblastoma cell U87MG compared with the DNA tetrahedron and the double-stranded DNA structures. We found that the DNA tetrahedron plays the main role in the endocytosis of U87MG cells, whereas the tumor-penetrating peptide could also bind to transmembrane glycoprotein neuropilin-1 and mediate the endocytosis of the p-TDN nanostructure. Moreover, given the high efficiency of the growth inhibitory effect of the p-TDN loading doxorubicin hydrochloride, the p-TDN distinguishes itself as a promising candidate as an effective delivery carrier.

Treatment of metastatic breast cancer by combination of chemotherapy and photothermal ablation using doxorubicin-loaded DNA wrapped gold nanorods.

Despite the exciting advances in cancer therapy over past decades, tumor metastasis remains the dominate reason for cancer-related mortality. In present work, DNA-wrapped gold nanorods with doxorubicin (DOX)-loading (GNR@DOX) were developed for treatment of metastatic breast cancer via a combination of chemotherapy and photothermal ablation. The GNR@DOX nanoparticles induced significant temperature elevation and DOX release upon irradiation with near infrared (NIR) light as shown in the test tube studies. It was found that GNR@DOX nanoparticles in combination with laser irradiation caused higher cytotoxicity than free DOX in 4T1 breast cancer cells. Animal experiment with an orthotropic 4T1 mammary tumor model demonstrated that GNR@DOX nanoplatform significantly reduced the growth of primary tumors and suppressed their lung metastasis. The Hematoxylin and Eosin (H&E) and immunohistochemistry (IHC) staining assays confirmed that the tumor growth inhibition and metastasis prevention of GNR@DOX nanoparticles were attributed to their abilities to induce cellular apoptosis/necrosis and ablate intratumoral blood vessels. All these results suggested a considerable potential of GNR@DOX nanoplatform for treatment of metastatic breast cancer.

Effect of dihydromyricetin on benzo[a]pyrene activation in rats.

OBJECTIVES: Flavanol dihydromyricetin (DHM) has been shown to counteract acute ethanol (EtOH) intoxication and reduce excessive EtOH consumption. Since this flavonoid is being considered for human use, the in vivo study of DHM interactions with the cytochrome P450 (CYP) multienzyme system in the respect of metabolic activation of a model food-born carcinogen, benzo[a]pyrene (BaP), is of high importance. Flavonoids of known properties, alpha-naphthoflavone (ANF) and beta-naphthoflavone (BNF) were included into the study to compare their and DHM effects on BaP-DNA adduct formation. METH0 DS: The flavonoids were administered by oral gavage either 72 hrs prior or simultaneously with a single dose of BaP to experimental rats. The expression of CYP1A1/2 enzymes was examined based on the enzymatic activity with a marker substrate, 7-ethoxyresorufin, and on Western blots. The nuclease P1 version of the 32P-postlabeling assay was used to detect and quantify covalent DNA adducts formed by BaP. RESULTS: Treatment of rats with a single dose of DHM or ANF prior to or simultaneously with BaP did not produce an increase in levels of CYP1A1 and in formation of BaP-DNA adducts in liver. BNF, a known inducer of CYP1A1, showed a synergistic effect on BaP-mediated CYP1A1 induction and BaP activation in liver. Contrary to that, in small intestine the stimulatory effect of BNF on both parameters was not detected. Animal pre-treatment with DHM or ANF before BaP administration resulted in a significant elevation of BaP-DNA adducts, namely in the distal part of small intestine, while the CYP1A1 mediated 7-ethoxyresorufin-O-deethylation (EROD) was decreased markedly. It is important to note that under all regimens of animal treatment, DHM or ANF produced the higher inhibitory effect on the BaP-DNA adduct formation and BaP-induced EROD activity of CYP1A1 when administered simultaneously than sequentially with BaP. Our data show that DHM or ANF did not enhance the BaP-activation leading to BaP-mediated genotoxicity (the formation of BaP-DNA adducts) in rat liver, however, in small intestine the pretreatment of rats with these flavonoids may enhance BaP genotoxicity. CONCLUSIONS: The data indicate that the intake of DHM prior to or simultaneously with the administration of BaP may increase the risk of a BaP-induced tumorigenesis in small intestine.

Pyrrolizidine alkaloid-derived DNA adducts as a common biological biomarker of pyrrolizidine alkaloid-induced tumorigenicity.

Pyrrolizidine alkaloid-containing plants are the most common poisonous plants affecting livestock, wildlife, and humans. The U.S. National Toxicology Program (NTP) classified riddelliine, a tumorigenic pyrrolizidine alkaloid, as "reasonably anticipated to be a human carcinogen" in the NTP 12th Report on Carcinogens in 2011. We previously determined that four DNA adducts were formed in rats dosed with riddelliine. The structures of the four DNA adducts were elucidated as (i) a pair of epimers of 7-hydroxy-9-(deoxyguanosin-N(2)-yl)dehydrosupinidine adducts (termed as DHP-dG-3 and DHP-dG-4) as the predominant adducts; and (ii) a pair of epimers of 7-hydroxy-9-(deoxyadenosin-N(6)-yl)dehydrosupinidine adducts (termed as DHP-dA-3 and DHP-dA-4 adducts). In this study, we selected a nontumorigenic pyrrolizidine alkaloid, platyphylliine, a pyrrolizidine alkaloid N-oxide, riddelliine N-oxide, and nine tumorigenic pyrrolizidine alkaloids (riddelliine, retrorsine, monocrotaline, lycopsamine, retronecine, lasiocarpine, heliotrine, clivorine, and senkirkine) for study in animals. Seven of the nine tumorigenic pyrrolizidine alkaloids, with the exception of lycopsamine and retronecine, are liver carcinogens. At 8-10 weeks of age, female F344 rats were orally gavaged for 3 consecutive days with 4.5 and 24 µmol/kg body weight test article in 0.5 mL of 10% DMSO in water. Twenty-four hours after the last dose, the rats were sacrificed, livers were removed, and liver DNA was isolated for DNA adduct analysis. DHP-dG-3, DHP-dG-4, DHP-dA-3, and DHP-dA-4 adducts were formed in the liver of rats treated with the individual seven hepatocarcinogenic pyrrolizidine alkaloids and riddelliine N-oxide. These DNA adducts were not formed in the liver of rats administered retronecine, the nontumorigenic pyrrolizidine alkaloid, platyphylliine, or vehicle control. These results indicate that this set of DNA adducts, DHP-dG-3, DHP-dG-4, DHP-dA-3, and DHP-dA-4, is a common biological biomarker of pyrrolizidine alkaloid-induced liver tumor formation. To date, this is the first finding that a set of exogenous DNA adducts are commonly formed from a series of tumorigenic xenobiotics.

A core cross-linked polymeric micellar platium(IV) prodrug with enhanced anticancer efficiency.

A core cross-linked polymeric micellar cisplatin(IV) conjugate prodrug is prepared by attaching the cisplatin(IV) to mPEG-b-PLL biodegradable copolymers to form micellar nanoparticles that can disintegrate to release the active anticancer agent cisplatin(II) in a mild reducing environment. Moreover, in vitro studies show that this cisplatin(IV) conjugate prodrug displays enhanced cytotoxicity against HepG2 cancer cells compared with cisplatin(II). Further studies demonstrate that the high cellular uptake and platinum-DNA adduct of this cisplatin(IV) conjugate prodrug can induce more cancer-cell apoptosis than cisplatin(II), which is responsible for its enhanced anticancer activity.

Alginate/CaCO3 hybrid nanoparticles for efficient codelivery of antitumor gene and drug.

In this study, a facile strategy for efficient codelivery of gene and drug was developed. Using a coprecipitation method, doxorubicin hydrochloride (DOX), an antitumor drug, and p53 expression plasmid were encapsulated in alginate/CaCO(3)/DNA/DOX nanoparticles with high encapsulation efficiency. The in vitro cell inhibition effect of the alginate/CaCO(3)/DNA/DOX nanoparticles was evaluated by MTT assay in HeLa cells. The alginate/CaCO(3)/DNA/DOX nanoparticles exhibited a high cell inhibition rate about 80%, indicating that the alginate/CaCO(3)/DNA/DOX nanoparticles could effectively mediate gene transfection and deliver the drug to the cells. Compared with the codelivery of gene and drug, the treatments by alginate/CaCO(3)/DOX nanoparticles and alginate/CaCO(3)/DNA nanoparticles separately led to much lower cell inhibition rates. Compared with the CaCO(3)/DNA/DOX nanoparticles without alginate modification, the alginate/CaCO(3)/DNA/DOX nanoparticles with a decreased particle size exhibited enhanced delivery efficiency. The alginate/CaCO(3)/DNA/DOX nanoparticles have promising applications in cancer treatments.

Delivery of interleukin-18 gene to lung cancer cells using cationic emulsion.

Interleukin-18 (IL-18) is known to reduce melanoma lung metastases through various mechanisms. For the delivery of IL-18 gene into the lung, three different cationic emulsions as non-viral vectors were formulated using the same components of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dioleoyl-sn-glycero-3-trimethylammonium propane (DOTAP), and Tween 80 with distinct oils. By using the small particle size of physicochemically stable E3, the complex of E3/plasmid DNA encoding IL-18 (16:2.5, w/w) was transfected into lung cancer cells, and the amount of plasmid DNA transferred and the expression of both mRNA and protein for IL-18 were measured. When compared with Lipofectamine/DNA complexes, an E3/DNA complex was less toxic and induced a comparable cellular level of plasmid DNA and expression levels of both mRNA and protein for IL-18. After injecting E3/DNA complexes into mice, the distribution of plasmid DNA was the highest in the lung and the liver. Especially, the administration of E3/DNA complexes induced a more rapid and prolonged distribution of plasmid DNA encoding IL-18 into the lung than that of Lipofectamine/DNA ones. These data demonstrated that cationic emulsion E3 containing castor oil could be useful for a delivery of IL-18 gene targeting the lung as well as the liver without an additional homing device, implying a potential IL-18 delivery system for the treatment of lung cancer.

[Chemotherapy of the small cell lung cancer using polyplatillen].

The experience of application of a new antitumoral platinum--containing preparation--polyplatyllen--in the treatment of the small cell pulmonary cancer was summarized. It was established, that the security profile of polyplatyllen is more favourable and efficacy in the treatment of the extended small cell pulmonary cancer is higher, than those of the standard scheme of treatment using cisplatinum and etoposide. The quality of life of the patients, while application of polyplatyllen, is more stable, than in application of the combined prescription of cisplatinum and etoposide.

[The survival indices of patients with stage IV gastric cancer treated by polyplatilen].

The survival indices of patients suffering gastric cancer stage IV in application of cytostatic preparation polyplatilen were studied up. The most expedient mode of treatment was suggested the conduction of no less than 2 courses of monochemotherapy. As a result, one year had survived (22.53 +/- 4.68)% patients, three years--(7.54 +/- 3.16)%, five years--(4.43 +/- 2.46)%.

[Results of palliative chemoradiotherapy of advanced small cell lung cancer using polyplatillen].

In the article authors summarize the experience of new anticancer drug polyplatillen usud in the chemoradiotherapy of small cell lung cancer. Polyplatillen improves the tumor response, general and relapse-free survival of advanced small cell lung cancer treatment in comparance with traditional VEC scheme.

[Improvement of the quality of life in patients with non-operable pancreatic cancer treated with polyplatillen].

The authors summarize in the article experience of a new anticancer drug Polyplatillen use in the chemotherapy of advanced pancreatic cancer. Polyplatillen use in the palliative chemotherapy in the patients with advanced pancreatic cancer permits to realize a control of pain and stabilize an ECOG status. Tendency to the improvement of total survival during an early period was registered in the group of patients with advanced pancreatic cancer who were treated with Polyplatillen.

[The efficacy of using low doses of immunomodulators for treating experimental tumors]. / Effektivnost' primeneniia malykh doz immunomoduliatorov dlia lecheniia éksperimental'nykh opukholei.

The data about the ability of various substances to cause definite reactions in biological objects are adduced, and, in particular, about the efficacy of a small doses of antitumoral chemopreparations in experiment. The results of investigation of the application efficacy of various doses of immunomodulating agents, such as arbisol, nucleoplat, the substrate from gram-negative bacteria Yersinia enterocolitica, antitumoral vaccine from the tumoral cells are presented for the cancer chemotherapy. While application of super-small doses of an antitumoral vaccine there was noted statistically significant inhibition of tumoral growth.

Mitoxantrone, etoposide and ara-C vs doxorubicin-DNA, ara-C, thioguanine, vincristine and prednisolone in the treatment of patients with acute myelocytic leukaemia. A randomized comparison.

Complex-binding of anthracyclines to DNA may increase their therapeutic efficacy. In a previous randomized trial patients with acute myelocytic leukaemia (AML) receiving combination chemotherapy including a DNA-bound doxorubicin preparation had a longer duration of first complete remission (CR) and survival than patients receiving free doxorubicin. In a parallel phase I/II study a combination of mitoxantrone, activity. In this randomized study of AML patients (15-60 years) induction treatment with MEA was compared to a combination of doxorubicin/DNA conjugate ara-C, thioguanine, vincristine and prednisolone (POCAL-DNA). The study was closed after an interim analysis of 86 patients. Thirty-five/42 (83%) and 20/44 (45%) patients entered CR in the MEA and POCAL-DNA groups, respectively (p < 0.001). With rescue therapy the corresponding figures were 88 and 64% (p < 0.02). Median survival was 27.8 and 13.1 months for MEA and POCAL-DNA patients, respectively (p < 0.03). In conclusion, the MEA regimen has a very high antileukaemic activity in good accordance with our previous experience. Since we could not reproduce our earlier clinical results using DNA-bound anthracyclines, the source and preparation of DNA seem to be of major importance.