Vinblastine and doxorubicin administration to pregnant mice affects brain development and behaviour in the offspring
Introduction
Yearly, more than 5 million children are born in Europe and it is estimated that 2500–5000 of these pregnancies are complicated by maternal cancer. The most frequently encountered malignancies are breast cancer and haematological malignancies, for which combination chemotherapy including doxorubicin and/or vinblastine, is the standard first line treatment (Cardonick and Iacobucci, 2004).
Cytotoxic drugs are designed to eradicate rapidly dividing cells, and thus, additionally to their effect on tumor cells, they may interfere with organogenesis and fetal growth (Xing et al., 1992, Wiebe and Sipila, 1994, Dawrant et al., 2007). Case reports are rare and mainly describe fetal losses, mutations and congenital malformations at birth, like skeletal and cardiac malformations, after the application of cytostatic drugs during early pregnancy (Zemlickis et al., 1993, Wiebe and Sipila, 1994, Ebert et al., 1997, Arnon et al., 2001, Cardonick and Iacobucci, 2004). Available data suggest a normal short-term outcome when chemotherapy is administered during the second or third trimester (Cardonick and Iacobucci, 2004, Ring et al., 2005, Hahn et al., 2006). However, the use of chemotherapeutics during the fetal phase of pregnancy might affect tissues like the nervous system, in which organogenesis proceeds well into the early postnatal period and defects of which will only become clear at a later age. Application of other toxins like heavy metals, ethanol and cocaine are known to adversely affect development during late pregnancy (Koren and Nulman, 1994). As chemotherapeutic drugs have a significant teratogenic potential far beyond the toxins listed above, their transplacental transport and impact on the developing fetus need to be characterized in detail. The more so since anatomical aspects of neural development of children prenatally exposed to chemotherapy have remained largely unstudied, and while data on functional aspects seem reassuring, analysis has been superficial so far (Ring et al., 2005, Hahn et al., 2006).
In adult rodents, doxorubicin reaches and damages neurohemal regions (neurohypophysis, median eminence, postremal area), but is largely excluded from brain regions with a functional blood–brain barrier (BBB) (Bigotte et al., 1982, Bigotte and Olsson, 1983, Hsieh et al., 2008). Circumventing the BBB by direct microinjection into the cerebral ventricles led to accumulation of the drug in neuronal and glial cell nuclei of the surrounding parenchyma (Bigotte and Olsson, 1984). As the BBB is not yet functional in the fetal and perinatal CNS, any drug that effectively passes the placenta may also reach the fetal CNS. Moreover, the fetal CNS offers an expanded “target list” for cytostatic drugs, as events like cell proliferation and migration, axon outgrowth and synaptogenesis are still well in progress in the late-fetal CNS. An exception may be the oligodendral myelin sheaths, which largely form after birth. In summary, the use of anthracyclins and vinca-alkaloids during pregnancy may cause a unique set of nervous system damage different from neurological side effects seen in children or adults.
Here we aimed to relate transplacental passage of vinblastine and doxorubicin to brain pathology and behavioural impairments in the offspring in a mouse model. Preliminary experiments revealed a potential impact on layer formation during fetal cortical development. Therefore, we have focused on cortical brain regions, i.e. cerebral (neo)cortex, hippocampus and cerebellum.
Drugs were administered on gestational day (GD) 17.5, after ending the embryonic development (Carnegie stage 23 is reached at GD 16.5) and corresponding to the peak of cell proliferation and migration in the mouse brain. Key events of neuron production, migration and settling occur during region-specific periods in mice, e.g. between GD 13 and P 2 in the murine neocortex (Hatten, 1999) or between GD 13 and about P 20 in cerebellum (Altman and Bayer, 1978) and hippocampal formation (Altman and Bayer, 1990, Stanfield and Cowan, 1979), when in the two latter cases the protracted proliferation of interneurons is taken into account. At GD 17.5, in all three major cortical brain regions cited above both the positioning of the respective macroneurons (hippocampal pyramidal neurons and subgranular neocortical pyramidal neurons, cerebellar Purkinje cells) and the proliferation of the microneurons is well underway. The use of the maximum tolerated dose at this stage should reveal effects both related to the final positioning of the cells defining the local architecture as well as effects related to cell proliferation and migration.
Both immediate (24 h p.i.) and residual (4–5 months p.i.) brain damage were investigated by light- and electron microscopy, and a battery of behavioural tests was performed in 3-month-old offspring.
Section snippets
Drug dosages
In the clinical setting, the most frequently applied dose of vinblastine is 6 mg/m2 per cycle, and of doxorubicin 25–60 mg/m2 corresponding to 0.2 mg/kg for vinblastine and 2 mg/kg for doxorubicin. Considering the inter-species differences in the metabolism and effect of drugs, for most drug dosages applied in mice should be much higher than in humans to reach the same effect (Collins, 2001). Compared to humans, mice are much less sensitive to vinca-alkaloids (van Tellingen et al., 1993). In
Transplacental transfer
Doxorubicin levels in fetal plasma reached 5.0 ± 0.2% of the maternal levels. For vinblastine a transplacental transfer of 13.9 ± 2.4% was measured (Table 1). In 1 out of 7 vinblastine-exposed mice, transfer was measured as 47%, which was much higher than the values obtained in the other 6 mice, despite an identical experimental procedure and sample processing and analysis. Therefore, we considered this value as an outlier.
General appearance
At GD 18.5, the frequency of visible fetal resorptions was comparable
Discussion
Currently, chemotherapy in pregnant women is postponed to the second and third trimester to avoid severe teratogenic effects on the fetus (Wiebe and Sipila, 1994, Ebert et al., 1997, Cardonick and Iacobucci, 2004, Dawrant et al., 2007). As this will basically shift the risk of damage to late-developing tissues like the nervous system, detailed assessment of brain development and resulting neurological performance is crucial for fine-tuning treatment regimens and to provide targeted diagnostic
Conclusions
Application of high dosages of doxorubicin and vinblastine during late gestation in mice causes transient regionally limited lesions to brain microvasculature and surrounding parenchyma and an inconsistent development of cortical ectopias resembling microgyri and isolated lissencephaly type 2-like overmigrations. The immediate lesions do not seem to leave permanent traces. Functional testing of the drug-exposed offspring indicated a few subtle, but specific alterations in their neurobehavioural
Conflict of interest statement
The authors declare that there are no conflicts of interest.
Acknowledgments
The research is financially supported by Research Foundation-Flanders (F.W.O.) Project G. 0358.06, ‘Stichting tegen kanker Project SCIE2006-17’, Research Fund-K.U. Leuven. Frédéric Amant is clinical researcher for Research Foundation-Flanders (F.W.O.); Kristel Van Calsteren is aspirant for Research Foundation-Flanders (F.W.O.).
References (52)
- et al.
Prenatal psychological stress causes higher emotionality, depression-like behavior, and elevated activity in the hypothalamo–pituitary–adrenal axis
Neurosci Res
(2007) - et al.
Hematological malignancies and pregnancy: a final report of 84 children who received chemotherapy in utero
Clin Lymphoma
(2001) - et al.
Multivariate neurocognitive and emotional profile of a mannosidosis murine model for therapy assessment
Neurobiol Dis
(2006) - et al.
Use of chemotherapy during human pregnancy
Lancet Oncol
(2004) Inter-species differences in drug properties
Chem Biol Interact
(2001)Exploratory behavior models of anxiety in mice
Neurosci Biobehav Rev
(1985)- et al.
Adriamycin produces a reproducible teratogenic model of vertebral, anal, cardiovascular, tracheal, esophageal, renal, and limb anomalies in the mouse
J Pediatr Surg
(2007) - et al.
Cytotoxic therapy and pregnancy
Pharmacol Ther
(1997) - et al.
Behavior ontogeny in the elevated plus-maze: prenatal stress effects
Int J Dev Neurosci
(2006) - et al.
Specific neurodevelopmental damage in mice offspring following maternal inflammation during pregnancy
Neuropharmacology
(2005)
Presenilin-1 deficiency leads to loss of Cajal-Retzius neurons and cortical dysplasia similar to human type 2 lissencephaly
Curr Biol
Effect of cyclosporin A on the brain regional distribution of doxorubicin in rats
Int J Pharm
Factors affecting grip strength testing
Neurotoxicol Teratol
Evaluation of marble-burying behavior as a model of anxiety
Pharmacol Biochem Behav
Antenatal maternal anxiety and stress and the neurobehavioural development of the fetus and child: links and possible mechanisms. A review
Neurosci Biobehav Rev
Pharmacology of antineoplastic agents in pregnancy
Crit Rev Oncol Hematol
The anti-cancer drug, doxorubicin, causes oxidant stress-induced endothelial dysfunction
Biochim Biophys Acta
Cognitive effects of standard-dose chemotherapy in patients with cancer
Cancer Invest
Prenatal development of the cerebellar system in the rat. I. Cytogenesis and histogenesis of the deep nuclei and the cortex of the cerebellum
J Comp Neurol
Migration and distribution of two populations of hippocampal granule cell precursors during the perinatal and postnatal periods
J Comp Neurol
Genetic and teratogenic effects of cancer treatments on gametes and embryos
Hum Reprod Update
F4/80, a monoclonal antibody directed specifically against the mouse macrophage
Eur J Immunol
Cytofluorescence localization of adriamycin in the nervous system. I. Distribution of the drug in the central nervous system of normal adult mice after intravenous injection
Acta Neuropathol
Toxic effects of adriamycin on the central nervous system. Ultrastructural changes in some circumventricular organs of the mouse after intravenous administration of the drug
Acta Neuropathol
Cytotoxic effects of adriamycin on the central nervous system of the mouse—cytofluorescence and electron-microscopic observations after various modes of administration
Acta Neurol Scand Suppl
Anxiety induced by prenatal stress is associated with suppression of hippocampal genes involved in synaptic function
J Neurochem
Cited by (18)
The impact of cancer and chemotherapy during pregnancy on child neurodevelopment: A multimodal neuroimaging analysis
2020, EClinicalMedicineCitation Excerpt :Moreover, the impact of chemotherapy on neurocognitive development remains a concern as the foetal brain is rapidly developing during the second and third trimesters of pregnancy through processes of neurogenesis, neuronal migration, synaptogenesis, etc. [5,6]. One mouse study observed prenatal exposure to vinblastine and doxorubicin to affect brain development, impacting both brain structure and behaviour [7]. Such early life impact might only become apparent in later life as the child develops into adolescence [8] and adulthood [5].
Increasing DNA binding affinity of doxorubicin by loading on Fe<inf>3</inf>O<inf>4</inf> nanoparticles: A multi-spectroscopic study
2020, Spectrochimica Acta - Part A: Molecular and Biomolecular SpectroscopyCitation Excerpt :DOX is a more common anti-cancer drug that used to chemotherapy of wide range of human cancers. Clinical studies revealed that intracranial injection of DOX specifically diminishes tumor volume [2]. The anticancer action of DOX is achieved by inhibiting topoisomerase II and free radical production, which leads to DNA damage and cell death [3,4].
Improving glioblastoma therapeutic outcomes via doxorubicin-loaded nanomicelles modified with borneol
2019, International Journal of PharmaceuticsCitation Excerpt :Doxorubicin (DOX) is a broad-spectrum anthracycline antitumor drug whose antitumor mechanism lies in inserting into the double helix strands of DNA, inhibiting topoisomerase II and then blocking DNA and RNA synthesis (Alrushaid et al., 2017). Clinical experimental studies have shown that the intracranial injection of doxorubicin via the internal carotid artery or vertebral artery has a significant therapeutic effect on glioma, improves clinical symptoms, and notably reduces tumor volume (Van Calsteren et al., 2009; Xu et al., 2017). However, doxorubicin does not effectively pass through the BBB and causes serious adverse reactions, especially cardiotoxicity (Khattry et al., 2009).
Management of cancer in pregnancy
2015, Best Practice and Research: Clinical Obstetrics and GynaecologyCitation Excerpt :For all investigated drugs, lower fetal concentrations were encountered. The transfer of chemotherapy is analyzed in animal models and in vitro [48–53], and it depends on maternal pharmacokinetics, placental blood flow, and the physicochemical drug properties [54]. The placenta is an active organ where placental transporters guide the transplacental passage of drugs.