A four-hour topotecan infusion achieves cytotoxic exposure throughout the neuraxis in the nonhuman primate model: implications for treatment of children with metastatic medulloblastoma.

The purpose of this study was to define the length of topotecan (TPT) i.v. infusion necessary to attain a cytotoxic exposure for medulloblastoma cells throughout the neuraxis. In vitro studies of human medulloblastoma cell lines (Daoy, SJ-Med3) were used to estimate the length and extent of TPT systemic exposure associated with inhibition of tumor cell growth or the exposure duration threshold (EDT). We evaluated TPT systemic and cerebrospinal fluid (CSF) disposition in six male rhesus monkeys (8-12 kg) that received TPT 2.0 mg/m2 i.v. as a 30-min or 4-h infusion. Plasma and CSF samples were assayed for TPT lactone by high-performance liquid chromatography, and the CSF exposures were compared with the estimated EDT. Results of the in vitro studies defined an EDT as a TPT lactone concentration of > 1 ng/ml for 8 h (IC99) daily for 5 days. The mean +/- SD for systemic clearance (CL(SYS)), penetration into fourth ventricle (%CSF(4th)), and penetration into lumbar space (%CSF(LUM)) were similar for the 30-min and the 4-h infusions. At a TPT lactone systemic exposure (AUC(PL)) of 56.7 +/- 19.9 ng/ml x h, time above 1 ng/ml in the fourth ventricle was 1.4-fold greater for a 4-h infusion compared with a 30-min infusion. At a TPT lactone AUC(PL) of 140 ng/ml x h, the 4-h infusion achieved the desired TPT exposure throughout the neuraxis (lateral and fourth ventricles and lumbar space), whereas the 30-min infusion failed to achieve it in the lumbar space. In conclusion, prolonging TPT i.v. infusion from 30-min to 4-h at a targeted AUC(PL) achieves the EDT throughout the neuraxis and represents an alternative method of TPT administration that will be tested prospectively in patients with high-risk medulloblastoma.

Delivery of adenoviral vectors to the prostate for gene therapy.

Prostate cancer has become the most frequently occurring cancer and the second leading cause of cancer deaths in men. One novel approach to combat prostate cancer is gene therapy. A replication-deficient recombinant adenoviral vector (AdRSVlacZ) expressing bacterial beta-galactosidase (beta-gal) (lacZ) under the control of the Rous sarcoma virus promoter was used to determine which delivery route was best for the transduction of adenoviral vectors to the prostate. Using a canine model, adenoviral vectors were administered by intravenous, intra-arterial, and intraprostatic (i.p.) injections. After injections, the expression of the lacZ gene was measured in canine prostates as well as in various other organs to determine the distribution of the disseminated adenoviral vector by (a) the percentage of cells expressing lacZ in situ (5-bromo-4-chloro-3-indolyl beta-D-galactoside staining), (b) beta-gal enzymatic activity (colorimetric beta-gal assay), and (c) polymerase chain reaction of genomic DNA using primers specific for the adenoviral genome. An i.p. injection of the adenoviral vector resulted in a greater transduction rate and expression level of lacZ in the prostate than either intravenous or intra-arterial (inferior vesical/prostatic artery) injections. Thus, an i.p. (or intratumoral) injection seems to be the best route to treat local regional prostate cancer by viral-based gene therapy.

The nature and action of host signals.

It is clear that excystations in vitro of the coccidia so far examined involves two steps, in the first of which CO2 is important, and the second, in which an external source of chymotrypsin and surface-active agents are required. However, the details of the mechanism of excystment are not clear. We do not know how the presence of CO2 changes the permeability of the oocyst wall. We do not know whether CO2 does anything to the sporozoite or sporocyst; the circumstance that mechanically-released sporocysts readily excyst under appropriate conditions without the necessity for high concentrations of, or perhaps any, CO2 suggests it does not. Circumstantial evidence suggests that the substrate in which chymotrypsin acts is the Stieda body, but whether the enzyme has other roles we do not know. Similarly, the role of bile is ill-defined, although it does seem that the induction of activity is important--but how is this brought about? The techniques available to excyst oocysts are, for many species, very efficient. If CO2 is, as it seems to be, a fundamental stimulus, then efficiency might be enhanced if more attention was given, not so much to increasing the time of exposure and amount of CO2 in the gas phase, but rather to the pH of the medium, which is rarely stated or apparently, controlled. The pH determines the proportion of the different carbonate species in solution, which may be of greater significance than the partial pressure of CO2 in the gas phase (see also Section V A). Although high numbers of excysted sporocysts can be obtained with a particular technique, this does not necessarily mean that all the signals supplied by the host are reproduced in vitro. Jackson (1962) found it necessary to wash oocysts in water or dilute buffers between the primary phase and the secondary phase, a step which implies a deficiency in the methods he used. Commonly, oocysts are exposed to a strong solution of L-cysteine. Does this reflect a general deficiency in the technique, or a counterpart of strongly reducing conditions in ruminant and non-ruminant alike? It seems that we have only a very general outline of excystment, and that we do not understand the details. Yet the problem seems to have been put aside; the most recent relevant reference we have found is dated 1983.