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Biomedical subjects

T Kottke

Publications and source records attributed to T Kottke.

14 recordsLinked to original sources
PubMed2006

VSV-G pseudotyped, MuLV-based, semi-replication-competent retrovirus for cancer treatment.

Low levels of gene delivery in vivo using replication-defective retroviral vectors have severely limited their application for clinical protocols. To overcome this problem, we describe here a semi-replication-competent retrovirus (s-RCR) in which the gag-pol and envelope (VSV-G, vesicular stomatitis virus G protein) genes were split into two vectors. This system offers potential advantages over both replication-defective vectors, in terms of efficiency of in vivo spread through a tumor, and all-in-one replication-competent vectors in terms of the payload of therapeutic genes that can be carried. We achieved a viral titer of s-RCR viruses approximately 70-fold higher than VSV-G pseudotyped, replication-defective vectors. In addition, s-RCR vectors induced tumor killing by the cytotoxicity of VSV-G during viral spread. Inclusion of the herpes simplex virus thymidine kinase (HSVtk30) gene into vectors significantly improved tumor killing activity followed by ganciclovir (GCV) treatment in vitro under conditions of low-level viral replication. However, at high levels of viral spread, VSV-G-mediated cytotoxicity predominated. Xenografts of human fibrosarcoma HT1080 cells, preinfected by semi-replicative green fluorescent protein vectors (semi-GFP), were completely non-tumorigenic in nude mice. Implantation of cells preinfected by semi-replicative TK30 vectors (semi-TK30) mixed with parental HT1080 cells at a ratio of 1:1 efficiently prevented tumor growth in mice treated by GCV. Direct intratumoral injection of HT1080 tumors growing in nude mice, or B16 murine melanoma in immunocompetent mice, with semi-TK30 viruses significantly prolonged survival. Injection of autologous cells (B16) producing semi-TK30 vector into B16 tumors prolonged survival only in mice treated with GCV but not with phosphate-buffered saline (PBS). In contrast, when xenogeneic cells (293T) producing semi-TK30 vectors were injected into B16 tumors, an optimal survival advantage was obtained in mice treated with PBS rather than GCV. These data indicate that complex interactions exist between direct cytotoxicity of VSV-G and HSVtk expression when placed in the context of additional immune parameters, which combine to determine the efficacy of the therapy. Taken together, our data suggest that s-RCR vectors have some potential advantages for development to deliver genes into tumors for cancer treatment but that a combination of factors will impact on the decision as to whether the s-RCR strategy is worth developing to full clinical trials.

Animals↗
PubMed2006

The perforin-dependent immunological synapse allows T-cell activation-dependent tumor targeting by MLV vector particles.

We have reported that retroviral particles adhered to the surface of antigen-specific T cells can be carried to metastases following adoptive transfer in vivo, a process we have called viral hitch hiking. Following antigen-driven T-cell accumulation at tumors, viral particles productively infect tumor cells via envelope/receptor dependent interactions ('hand on' of virus from the T cell to the tumor cell). We describe here a second envelope/receptor independent pathway of viral hand on from T cells, dependent on T-cell activation. We show that the endosomolytic property of perforin promotes release of viral particles from endosomes into which they are co-delivered along with cytotoxic granules from the activated T cell. Therefore, hand on of MLV particles lacking any envelope can be used for in vivo delivery of vectors, where targeting is at the extremely specific level of recognition of antigen by the T-cell receptor, thereby dispensing with the need to engineer viral envelopes. These data reveal a novel pathway by which MLV viral particles exploit a functional immunological synapse and present new opportunities both to improve the efficacy of adoptive T-cell transfer and to target vectors for systemic gene delivery.

Adoptive Transfer↗
PubMed2003

Intratumoral expression of a fusogenic membrane glycoprotein enhances the efficacy of replicating adenovirus therapy.

We describe here a novel strategy to enhance the in vivo efficacy of replicating adenovirus therapy, using coinjection of plasmid DNA encoding a fusogenic viral glycoprotein. The combination of fusogenic membrane glycoprotein (FMG)-induced tumor cell fusion and infection with replicating adenovirus effectively treats even large established tumors at doses of plasmid DNA and virus that alone are ineffective. Adenoviral infection appears to increase the transduction of the tumor cells to a modest degree thereby boosting the FMG-mediated component of the therapy. Simultaneously, syncytial formation enhances the therapeutic effects of viral infection by increasing spread of adenoviral particles through the tumor cell population and by increasing titer of virus released from the tumor cells. This effect is due probably to release of intracellular viral particles upon tumor cell death and also to increased levels of E1A protein within syncytia, whose increased metabolic rate is associated with enhanced levels of protein expression. Cotransduction of tumor cells with replicating adenovirus and FMG-expressing vectors could either be combined within single replicating vectors or could be used in strategies using separate administration of two components, both at lower doses than required for either therapy alone.

Adenoviridae↗
PubMed2001

A community-wide smoking cessation program: Quit and Win 1998 in Olmsted county.

BACKGROUND: Quit and Win is a community-wide stop smoking contest to help cigarette smokers stop smoking and educate the general public concerning smoking hazards. METHODS: All community residents, 15 years of age or older, were eligible to participate in either the stop smoking contest or the supporter contest. A random telephone survey to local households was conducted before and after the Quit and Win contest to assess the level of knowledge and attitude changes about smoking. RESULTS: Of the 304 smokers enrolled in the contest, 42% self-reported continuous tobacco abstinence for the 4-week contest period and 11% were abstinent at 1 year postcontest. Significant predictors for tobacco abstinence during the contest were formal education beyond high school, absence of other smokers in the household, having a support person enrolled in the support person contest, and the type of relationship that the support person had with their smoker. Survey results showed that this contest changed some local attitudes and increased general knowledge of smoking hazards. CONCLUSIONS: Community-wide stop smoking contests can be used to engage smokers and their support in the community and can be successful in reducing tobacco use.

Adolescent↗
PubMed1999

Caspase-mediated cleavage of DNA topoisomerase I at unconventional sites during apoptosis.

Previous studies have demonstrated that topoisomerase I is cleaved late during apoptosis, but have not identified the proteases responsible or examined the functional consequences of this cleavage. Here, we have shown that treatment of purified topoisomerase I with caspase-3 resulted in cleavage at DDVD146 downward arrowY and EEED170 downward arrowG, whereas treatment with caspase-6 resulted in cleavage at PEDD123 downward arrowG and EEED170 downward arrowG. After treatment of Jurkat T lymphocytic leukemia cells with anti-Fas antibody or A549 lung cancer cells with topotecan, etoposide, or paclitaxel, the topoisomerase I fragment comigrated with the product that resulted from caspase-3 cleavage at DDVD146 downward arrowY. In contrast, two discrete topoisomerase I fragments that appeared to result from cleavage at DDVD146 downward arrowY and EEED170 downward arrowG were observed after treatment of MDA-MB-468 breast cancer cells with paclitaxel. Topoisomerase I cleavage did not occur in apoptotic MCF-7 cells, which lack caspase-3. Cell fractionation and band depletion studies with the topoisomerase I poison topotecan revealed that the topoisomerase I fragment remains in proximity to the chromatin and retains the ability to bind to and cleave DNA. These observations indicate that topoisomerase I is a substrate of caspase-3 and possibly caspase-6, but is cleaved at sequences that differ from those ordinarily preferred by these enzymes, thereby providing a potential explanation why topoisomerase I cleavage lags behind that of classical caspase substrates such as poly(ADP-ribose) polymerase and lamin B1.

Animals↗
PubMed1997

Activation of multiple interleukin-1beta converting enzyme homologues in cytosol and nuclei of HL-60 cells during etoposide-induced apoptosis.

Recent genetic and biochemical studies have implicated cysteine-dependent aspartate-directed proteases (caspases) in the active phase of apoptosis. In the present study, three complementary techniques were utilized to follow caspase activation during the course of etoposide-induced apoptosis in HL-60 human leukemia cells. Immunoblotting revealed that levels of procaspase-2 did not change during etoposide-induced apoptosis, whereas levels of procaspase-3 diminished markedly 2-3 h after etoposide addition. At the same time, cytosolic peptidase activities that cleaved DEVD-aminotrifluoromethylcoumarin and VEID-aminomethylcoumarin increased 100- and 20-fold, respectively; but there was only a 1. 5-fold increase in YVAD-aminotrifluoromethylcoumarin cleavage activity. Affinity labeling with N-(Nalpha-benzyloxycarbonylglutamyl-Nepsilon-biotin yllysyl)aspartic acid [(2,6-dimethylbenzoyl)oxy]methyl ketone indicated that multiple active caspase species sequentially appeared in the cytosol during the first 6 h after the addition of etoposide. Analysis on one- and two-dimensional gels revealed that two species comigrated with caspase-6 and three comigrated with active caspase-3 species, suggesting that several splice or modification variants of these enzymes are active during apoptosis. Polypeptides that comigrate with the cytosolic caspases were also labeled in nuclei of apoptotic HL-60 cells. These results not only indicate that etoposide-induced apoptosis in HL-60 cells is accompanied by the selective activation of multiple caspases in cytosol and nuclei, but also suggest that other caspase precursors such as procaspase-2 are present but not activated during apoptosis.

Antineoplastic Agents, Phytogenic↗
PubMed1997

The multilevel compliance challenge: recommendations for a call to action. A statement for healthcare professionals.

Despite the universally accepted importance of compliance, strategies known for more than two decades to be effective are not routinely incorporated into clinical practice. For the benefits of primary and secondary prevention to be realized in diverse population groups and settings, emphasis must be placed on implementing strategies at the patient, provider, and organization levels. Current knowledge of compliance strategies, if integrated into a multilevel approach, offers enormous promise for decreasing risk and improving patient outcomes.

American Heart Association↗
PubMed1992

Body mass index and the initial manifestation of coronary heart disease in women aged 40-59 years.

A population-based case-control study was conducted to evaluate body mass as a coronary heart disease (CHD) risk factor among women 40-59 years of age. Cases were women aged 40-59 whose first manifestation of CHD was angina (No. = 133), nonfatal myocardial infarction (No. = 90), and sudden unexpected death (No. = 18) during 1960-1982. Two randomly selected controls were matched on age and time of the initial disease manifestation of the case. The adjusted relative risk for weight and body mass index respectively demonstrated a moderate association with all CHD as well as with angina, but no association with definite CHD (myocardial infarction or sudden unexpected death). To determine if the observed association between body mass index and angina was possibly attributable to differential misclassification bias (i.e. obese women were, in contrast to non-obese women, preferentially labelled as having coronary artery disease) data for angina were stratified by confirmed versus unconfirmed cardiac origin. In the unconfirmed angina analysis, the 75th percentile for weight contrasted with the 25th percentile was associated with a 50% increase in the risk of being labelled as having angina (adjusted odds ratio (OR) = 1.59, 95% confidence interval (CI): 1.11-2.28), while a similar contrast for Quetelet Index was also associated with a nearly 2-fold increase in the risk of being labelled as having angina (adjusted OR = 1.74, 95% CI: 1.18-2.57).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1991

Polychotomous multivariate models for coronary heart disease simulation. I. Tests of a logistic model.

Stochastic compartmental modeling techniques have been employed to simulate coronary heart disease morbidity and mortality. In the current paper, polychotomous logistic models are used to describe the relationship between risk of disease and multiple risk factors, effect modification and confounding variables. The process of estimating the parameters for two risk factors and three types of outcomes is described for a population followed for five years. A Statistical Analysis System (SAS) procedure was used to estimate risk factor coefficients based on two partial periods and on the entire five year epoch. Most of the estimated coefficients were found to be statistically significant. The model performance was evaluated by comparing the observational data with simulated outcomes using a micropopulation and Monte Carlo techniques. Two different tests of goodness of fit were used. Satisfactory fits were obtained both for the risk coefficients based on two partial periods and those based on the entire epoch. This indicates that the model is suitable for simulation of the effects of intervention strategies. The use of the entire epoch involved estimates of one half as many parameters as did the use of two partial periods. Accordingly, it is concluded that only the entire epoch need be considered for future studies of this population.

Adult↗
PubMedDate not supplied

Polychotomous multivariate models for coronary heart disease simulation. II. Comparisons of risk functions.

This is the second in a series of papers dealing with models of coronary heart disease. Three different types of statistical models are considered as risk functions: the multivariate logistic model, the Cox proportional hazard model and the Neyman exponential risk avoidance model. The types of models differ in the form hypothesized for the probability of occurrence of coronary heart disease outcomes: incident myocardial infarct, cardiac death, and death from other causes. Although the three risk functions are strikingly different, they can all be tested using the CRISPERS chronic disease simulation system. Simulations were performed using data from North Karelia, Finland. The polychotomous multivariate logistic risk function is convenient for studies involving increasing numbers of risk factors. The Cox proportional hazard regression model is shown to be unsuitable for the cohort dataset used as well as for some of the intended uses of the simulation models. The Neyman exponential risk avoidance model involves time in a quite different fashion. It has the inherent advantage of being easier to relate to underlying biological mechanisms because it is the integral of first order rate equations. It is concluded that more than one risk function should be evaluated for simulations of coronary heart disease.

Adult↗
PubMedDate not supplied

Polychotomous multivariate models for coronary heart disease simulation. III. Model sensitivities and risk factor interventions.

This is the third in a series of papers dealing with models of coronary heart disease. Sensitivity analyses of the logistic risk function and the Neyman risk function are reported. The resulting response surfaces are also used to investigate the optimality of the set of values for the risk coefficients. It is shown that the coefficients estimated by maximum likelihood are preferable to the sets from an optimisation procedure. Two different sets of risk coefficients estimated using short periods and entire epochs for the logistic risk function are shown to lead to similar conclusions concerning simulated primary intervention strategies. However, the corresponding risk factor reductions using the Neyman risk function lead to somewhat different effects. Additional information is needed to distinguish between these two assumptions of the risk function used to model coronary heart disease. This underscores the need to understand the effects of the underlying risk function assumed when interpreting simulated outcomes of intervention strategies.

Computer Simulation↗