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Auger radiation targeted into DNA: a therapy perspective.

BACKGROUND: Auger electron emitters that can be targeted into DNA of tumour cells represent an attractive systemic radiation therapy goal. In the situation of DNA-associated decay, the high linear energy transfer (LET) of Auger electrons gives a high relative biological efficacy similar to that of alpha particles. In contrast to alpha radiation, however, Auger radiation is of low toxicity when decaying outside the cell nucleus, as in cytoplasm or outside cells during blood transport. The challenge for such therapies is the requirement to target a high percentage of all cancer cells. An overview of Auger radiation therapy approaches of the past decade shows several research directions and various targeting vehicles. The latter include hormones, peptides, halogenated nucleotides, oligonucleotides and internalising antibodies. DISCUSSION: Here, we will discuss the basic principles of Auger electron therapy as compared with vector-guided alpha and beta radiation. We also review some radioprotection issues and briefly present the main advantages and disadvantages of the different targeting modalities that are under investigation.

DNA Damage↗

Radioimmunotherapy with alpha-emitting nuclides.

This review discusses the application of alpha particle-emitting radionuclides in targeted radioimmunotherapy. It will outline the production and chemistry of astatine-211, bismuth-212, lead-212, actinium-225, bismuth-213, fermium-255, radium-223 and terbium-149, which at present are the most promising alpha-emitting isotopes available for human clinical use. The selective cytotoxicity offered by alpha particle-emitting radioimmunoconstructs is due to the high linear energy transfer and short particle path length of these radionuclides. Based upon the pharmacokinetics of alpha particle-emitting radioimmunoconstructs, both stochastic and conventional dosimetric methodology is discussed, as is the preclinical and initial clinical use of these radionuclides conjugated to monoclonal antibodies for the treatment of human neoplasia.

Alpha Particles↗

Thorotrast-induced cholangiocarcinoma: case report.

Thorotrast, a contrast medium used extensively before being banned in 1950s, delivers a densely ionizing, high linear energy transfer type of radiation that predisposes to malignancies. We report a case of peripheral cholangiocarcinoma and describe its computed tomographic and magnetic resonance imaging features in a patient who developed it 48 years after exposure to Thorotrast.

Aged↗

Relative effectiveness of different particles and energies in disrupting behavioral performance.

On exploratory class missions to other planets, astronauts will be exposed to varieties and doses of heavy particles, which are not experienced in low earth orbit. These particles can affect neurobehavioral function and potentially interfere with the ability of astronauts to successfully meet mission requirements. While a significant amount of research has been performed on the relative biological effectiveness (RBE) of different types of heavy particles on cytogenetic function, little research has been done on the effectiveness of different particles on central nervous system function and on cognitive/behavioral performance. The present paper reviews some recent research on the effects of exposure to different types and energies of heavy particles on the performance of two behavioral tasks which depend upon the integrity of the central dopaminergic system. This review indicates that the RBE of different particles for neurobehavioral dysfunction cannot be predicted only on the basis of the linear energy transfer of the specific particle.

Animals↗

Radial distribution of electron spectra from high-energy ions.

The average track model describes the response of physical and biological systems using radial dose distribution as the key physical descriptor. We report on an extension of this model to describe the average distribution of electron spectra as a function of radial distance from an ion. We present calculations of these spectra for ions of identical linear energy transfer (LET), but dissimilar charge and velocity to evaluate the differences in electron spectra from these ions. To illustrate the usefulness of the radial electron spectra for describing effects that are not described by electron dose, we consider the evaluation of the indirect events in microdosimetric distributions for ions. We show that folding our average electron spectra model with experimentally determined frequency distributions for photons or electrons provides a good representation of radial event spectra from high-energy ions in 0.5-2 microm sites.

Dose-Response Relationship, Radiation↗

Quantitative modelling of DNA damage using Monte Carlo track structure method.

This paper presents data on modelling of DNA damage induced by electrons, protons and alpha-particles to provide an insight into factors which determine the biological effectiveness of radiations of high and low linear energy transfer (LET). These data include the yield of single- and double-strand breaks (ssb, dsb) and base damage in a cellular environment. We obtain a ratio of 4-15 for ssb:dsb for solid and cellular DNA and a preliminary ratio of about 2 for base damage to strand breakage. Data are also given on specific characteristics of damage at the DNA level in the form of clustered damage of varying complexity, that challenge the repair processes and if not processed adequately could lead to the observed biological effects. It is shown that nearly 30% of dsb are of complex form for low-LET radiation, solely by virtue of additional breaks, rising to about 70% for high-LET radiation. Inclusion of base damage increases the complex proportion to about 60% and 90% for low- and high-LET radiation, respectively. The data show a twofold increase in frequencies of complex dsb from low-LET radiation when base damage is taken into account. It is shown that most ssb induced by high-LET radiation have associated base damages, and also a substantial proportion is induced by low-energy electrons.

DNA↗

The track structures of ionizing particles and their application to radiation biophysics. I. A new analytical method for investigating two biophysical models.

A new approach to the interpretation of the effects of radiation on cells is described, in which sample particle tracks are constructed using a Monte Carlo computer program and the exposure of cellular targets to these tracks is simulated using a second program known as BIOPHYS. Data on the shapes and DNA contents of the cell nuclei are obtained from the literature. It is assumed that the sensitive material is DNA, and that the target is divided into cubes of approximately 2 nm (the diameter of the DNA helix) per side; the numbers of these cubes containing different numbers of ionizations are derived. Two different methods of analysing the output of BIOPHYS are described. In the first, it is assumed that lethality is caused by the occurrence of a number of ionizations equal to or greater than a certain threshold in one cube; in the second method, it is assumed that only two ionizations are required, in different parts of the cube, but that only some fraction of the cube is sensitive. These models have been applied to the interpretation of the variation of radiosensitivity with a linear energy transfer (LET) of spores of Bacillus subtilis exposed wet and dry, and good fits to the published experimental data were obtained using both models. Fits to experimental data for a range of other cell lines will be presented in a second paper.

Bacillus subtilis↗

The efficacy of conventional external beam, three-dimensional conformal, intensity-modulated, particle beam radiation, and brachytherapy for localized prostate cancer.

Technologic advances in radiation treatment planning and delivery have generated popular interest in the different radiation therapy techniques used in treating patients with localized prostate cancer. Throughout the past decade, high-energy (> 4 MV) linear accelerators have largely replaced Cobalt machines in external beam radiation therapy (EBRT) delivery. Conventional EBRT has been used to treat prostate cancer successfully since the 1950s. By switching to computed tomography-based planning, three-dimensional conformal radiation therapy provides better relative conformality of dose than does conventional EBRT. Intensity-modulated radiation therapy (IMRT) has further refined dose conformality by spreading the low-dose region to a larger volume. However, the potential long-term risks of larger volumes of normal tissues receiving low doses of radiation in IMRT are unknown. Particle-beam radiation therapy offers unique dose distributions and characteristics with higher relative biologic effect and linear energy transfer. Transperineal prostate brachytherapy offers the shortest treatment time with equivalent efficacy without significant risk of radiation exposure. The addition of hormonal therapy to radiation therapy has been shown to improve the outcome of radiation therapy.

Antineoplastic Agents, Hormonal↗

The development of alpha-emitting radionuclide lead 212 for the potential treatment of ovarian carcinoma.

alpha-Emitting radionuclides may be an effective alternative treatment against ovarian carcinoma because they have short half-lives and are densely ionizing, with high linear energy transfer to a depth of several cell diameters without requiring cellular oxygenation. One radionuclide that has been generated and tested in our laboratory in vitro and in vivo is lead 212 (212Pb). Intraperitoneal instillation of 212Pb prolonged survival and totally eradicated tumor in 24% of mice inoculated with the extremely virulent Ehrlich ascites-producing tumor. In vitro 212Pb was two to four times more effective in killing human ovarian cancer cells than x-rays. Irradiation with 212Pb increased the radiosensitivity and chromosomal aberrations of cells. In dogs, intraperitoneal instillation of 2.6 mCi of ferrous hydroxide tagged with 212Pb caused no significant toxicity. It appears that alpha-emitting radionucides such as 212Pb have the potential to be a new and potent treatment of ovarian carcinoma and could be effective in cases that are resistant to conventional chemotherapy or x-ray therapy.

Animals↗

Intracellular distribution of various boron compounds for use in boron neutron capture therapy.

The neutron capture reaction in boron (10B(n, alpha)7Li) generates two short-range particles with high linear energy transfer. The effect of neutron capture therapy depends on the selective localization of 10B atoms in target cells. The determination of the distribution of boron compounds in cancer cells at the subcellular level is required for the understanding of the effect of this treatment. The monomeric sulfhydryl borane (BSH) compound has been used clinically in Japan and preclinically in the U.S.A. Recently, new compounds have been developed: a dimeric sulfhydryl borane (BSSB), a boronophenylalanine (BPA), and two porphyrin complexes (BOPP and VCDP). This study demonstrates that the porphyrin complexes (BOPP and VCDP) are more cytotoxic than the other three compounds to the rat 9L gliosarcoma cell line. Using atomic absorption spectrophotometry to determine boron content for cellular uptake studies of these agents, we found that of the five compounds tested BOPP (25 microM) exposure resulted in the greatest boron uptake averaging 305 ng B/10(6) cells. BSSB (500 microM) was second averaging 93 ng B/10(6) cells, BSH (500 microM) third averaging 62 ng B/10(6) cells, VCDP (25 microM) fourth averaging 58 ng B/10(6) cells, and BPA (500 microM) fifth averaging 7.4 ng B/10(6) cells. Data on the distribution of boron in the nuclei, mitochondria, lysosomes, microsomes, and cytosomes of 9L cells are also presented.

Animals↗

Relative effectiveness of neutrons and X-rays in induction of crossing over in Drosophila males.

Relative biological effectiveness of neutrons vs. X-rays in inducing crossing-over in males of D. melanogaster was investigated using 812 and 834 rad of neutrons and the same dose of X-rays. Crossing-over was induced in spermatocytes and spermatogonia of adults and pupae. Neutrons were 4 times more effective in spermatocytes of adults and their effectiveness in pupal spermatocytes was even more. Neutrons also induced more exchanges in spermatogonial cells including predefinitive spermatogonia. Higher effectiveness of neutrons can be attributed to their high linear energy transfer.

Animals↗

The study of post-irradiation recovery of yeast: the 'premolecular period'.

Recovery of yeast cells after exposure to ionizing radiation was discovered in 1957. During the following decade, i.e., in the 'premolecular period' of studying this phenomenon, its basic features including dependence on cell ploidy, cellular energy exchange and radiation linear energy transfer (LET) were determined and a mathematical model of recovery was proposed. In addition, DNA double-strand breaks were shown to be the damage causing death of irradiated haploid and diploid cells. The results of these investigations led to the formulation of the concept that the ability of a cell to repair genetic damage was of universal biological importance.

Cell Death↗

Changing patterns of radiosensitivity of hematopoietic progenitors from chronically irradiated dogs prone either to aplastic anemia or to myeloproliferative disease.

Hematopoietic patterns have been assessed in chronic 60Co gamma irradiated dogs during preclinical phases of evolving aplastic anemia (AA) or myeloproliferative disease (MPD), principally myeloid leukemia. Within the AA-prone dog, a singular phase of progressive decline in blood levels of granulocytes and monocytes was noted along with a similar reduction in marrow progenitors committed to granulocyte/monocyte differentiation (CFU-GM). Measured radioresistance of the preAA CFU-GM in vitro, relative to control CFU-GM from nonirradiated animals, revealed only slightly increased resistance to gamma rays, but significantly increased resistance to fission neutrons. Within the MPD-prone dogs, four preclinical phases (i.e. suppression, partial recovery, accommodation, and preleukemic transition) preceding development of overt MPD were evidenced by the monitored change in blood granulocyte/monocyte counts and marrow progenitor levels. Analysis of radioresistance of preMPD CFU-GM revealed marked changes with time of exposure and, in turn, with preclinical phase transitions. Gamma ray resistance increased in the initial phases of exposure, with maximal levels occurring during the middle phase of exposure (accommodation, phase III) followed by a tailing off of resistance at later times. Resistance to fission neutrons by preMPD CFU-GM was observed as well, but somewhat later in the exposure course and at a much lower, more consistent level. These differential patterns of radioresistance expressed by marrow CFU-GM of chronically irradiated MPD-prone dogs to gamma rays and fission neutrons gave rise to preclinical phase-specific 'relative biological effectiveness' (RBE) values. From these observations, we conclude that: (i) CFU-GM of MPD-prone dogs acquire and maintain marked radioresistance to low linear energy transfer (LET) gamma rays, but only marginally elevated radioresistance to high-LET fission neutrons during the course of chronic gamma ray exposure; and (ii) CFU-GM of the AA-prone dog, in contrast, acquire little change in resistance to gamma rays, but, surprisingly, marked resistance to neutrons relative to progenitors from nonirradiated controls. These results support the concept that acquired radioresistance of vital granulocyte/monocyte lineage-committed hematopoietic progenitors is temporally, perhaps causally, linked to the processes mediating hematopoietic recovery and accommodation under chronic irradiation, and in turn to preclinical events of evolving MPD. In addition, the marked differential responses of progenitors to gamma and neutron irradiation in vitro might suggest differences in the nature of cellular lesions elicited by chronic gamma irradiation, in vivo.

Anemia, Aplastic↗

Melphalan resistant human ovarian tumour cells are cross-resistant to photons, but not to high LET neutrons.

High linear energy transfer (LET) fast neutrons for the local control of advanced head and neck tumours are currently being evaluated at several centres. Fast neutrons are believed to produce more direct, and less OH mediated damage than photons, and consequently be less affected by intracellular thiol levels. Chemoresistant tumours with elevated thiol levels may therefore be more effectively controlled by fast neutron therapy than by photons. The "in vitro" radiation response of melphalan sensitive and resistant human ovarian tumour cell lines has demonstrated that melphalan resistance confers a 1.5-fold level of cross-resistance to photons, primarily attributable to a 2-fold decrease in the alpha component in the resistant OAW42/MER cell line. Pretreatment of the melphalan-resistant line with the thiol depleting agent buthionine sulphoximine (BSO) restored the magnitude of alpha to a value similar to that in the chemosensitive cell line. The survival curves of these cell lines following neutron irradiation were near exponential, with similar values of alpha. This study has demonstrated that melphalan resistant tumour cells are cross-resistant to photon irradiation, but not to fast neutrons. The mechanism of cross-resistance has yet to be determined, but glutathione (GSH) appears to be involved.

Fast Neutrons↗

Boron neutron capture irradiation of the rat spinal cord: effects of variable doses of borocaptate sodium.

The Fischer 344 rat spinal cord model has been used to evaluate the response of the central nervous system to boron neutron capture irradiation with variable doses of the neutron capture agent, borocaptate sodium (BSH). Three doses of BSH, 190, 140 and 80 mg/kg body weight, administered by i.p. injection, were used to establish the time course of 10B accumulation in and removal from the blood. After administration of the two lower doses of BSH, blood 10B levels peaked at 0.5 h after injection, with no significant (P > 0.1) change at 1 h after injection. Beyond this time point, levels of 10B in the blood began to decrease after a dose of 80 mg/kg BSH, but remained constant until 3 h after administration after the two higher doses of BSH. Myelopathy developed after latent intervals of 20.4 +/- 0.1, 20.8 +/- 1.4, 15.0 +/- 0.8, 15.4 +/- 0.4 and 15.6 +/- 0.4 weeks, following irradiation with thermal neutrons in combination with BSH at doses of 20, 40, 80, 140 and 190 mg/kg body weight, respectively. The radiation-induced lesion in the spinal cord was white matter necrosis. ED50 values for myelopathy were calculated from probit-fitted dose-effect curves. Expressed as total physical absorbed doses, these values were 20.7 +/- 1.9, 24.9 +/- 1.2, 27.2 +/- 0.9, 28.4 +/- 0.6 and 32.4 +/- 1.9 Gy after irradiation with thermal neutrons in the presence of 20, 40, 80, 140 and 190 mg/kg body weight of BSH, respectively. The compound biological effectiveness (CBE) factor values, estimated from this data, were in the range 0.49-0.55. There was no significant (P > 0.1) variation in the CBE factor for BSH as a function of increasing 10B concentration in the blood. It was concluded that there was no significant synergistic interaction between the low and high linear energy transfer (LET) components of the boron neutron capture (BNC) radiation field.

Animals↗

Effect of HZE particles and space hadrons on bacteriophages.

The effect of high energy (HZE) particles and high energy hadrons on T4Br+ bacteriophage was analyzed. The experiments were done in orbital flight, on high mountains, on an accelerator, and with an alpha particle source. We studied the survival rate of the bacteriophage, the mutation frequency, the mutation spectrum and the revertability under the action of chemical mutagens with a known mechanism of action on DNA. It was found that the biological efficiency of HZE particles and high energy hadrons is greater than that of gamma radiation. The spectra of mutations produced by these mutations and the mechanisms of their action are also different. These effects were local, because of the mode of interaction of the radiant energy with biological objects, and depended on the linear energy transfer (LET). The modes have now been experimentally defined.

Alpha Particles↗

Late skin damage in rabbits and monkeys after exposure to particulate radiations.

Skin biopsies were taken from the central regions of the ears of New Zealand white rabbits following localized exposure of one ear of each rabbit to 530 MeV/amu Ar or 365 MeV/amu Ne ions. The unirradiated ears served as controls. Biopsies were taken also from the chests and inner thighs of rhesus monkeys after whole-body exposure to 32 MeV protons and from unirradiated control animals. The linear energy transfers (LET infinity's) for the radiations were 90 +/- 5, 35 +/- 3, and approximately 1.2 keV/micrometer, respectively. In the rabbit studies, explants were removed with a 2 mm diameter dermal punch at post-irradiation times up to five years after exposure. Similar volumes of monkey tissue were taken from skin samples excised surgically 16-18 years following proton irradiation. Fibroblast cultures were initiated from the explants and were propagated in vitro until terminal senescence (cessation of cell division) occurred. Cultures from irradiated tissue exhibited decreases in doubling potential that were dependent on radiation dose and LET infinity and seemed to reflect damage to stem cell populations. The implications of these results for astronauts exposed to heavy ions and/or protons in space include possible manifestations of residual effects in the skin many years after exposure (e.g. unsatisfactory responses to trauma or surgery).

Age Factors↗

The measured radiation environment within Spacelabs 1 and 2 and comparison with predictions.

To measure the radiation environment in the Spacelab (SL) module and on the pallet, a set of passive and active radiation detectors was flown as part of the Verification Flight Instrumentation (VFI). SL 1 carried 4 passive and 2 active detector packages which, with the data from the 26 passive detectors of Experiment INS006, provided a comprehensive survey of the radiation environment within the spacecraft. SL 2 carried 2 passive VFI units on the pallet. Thermoluminescent dosimeters (TLDs) measured the low linear energy transfer (LET) dose component; the HZE fluence and LET spectra were mapped with CR-39 track detectors; thermal and epithermal neutrons were measured with the use of fission foils; metal samples analyzed by gamma ray spectroscopy measured low levels of several activation lines. The TLDs registered from 97 to 143 mrad in the SL 1 module. Dose equivalents of 330 +/- 70 mrem in the SL 1 module and 537 +/- 37 mrem on the SL 2 pallet were measured. The active units in the SL 1 module each contained an integrating tissue-equivalent ion chamber and two differently-shielded xenon-filled proportional counters. The ion chambers accumulated 125 and 128 mrads for the mission with 17 and 12 mrads accumulated during passages through the South Atlantic Anomaly (SAA). The proportional counter rates (approximately 1 cps at sea level) were approximately 100 cps in the middle of the SAA (mostly protons), approximately 35 cps at large geomagnetic latitudes (cosmic rays) and approximately 100 cps in the South Horn of the electron belts (mostly bremsstrahlung). Detailed results of the measurements and comparison with calculated values are described.

Atlantic Ocean↗