Placental, Foetal, and Maternal Serum Metabolomic Profiles in Pregnancy-Associated Cancer: Walker-256 Tumour Model in a Time-Course Analysis

Abstract

Cancer during pregnancy presents a delicate coexistence, imposing ethical and professional challenges on both the patient and medical team. In this study, we aimed to explore in a pre-clinical model the impact of tumour evolution in serum, placental and foetal metabolomics profiles during pregnancy in a time-course manner. Pregnant Wistar rats were distributed into two experimental groups: Control (C) and Walker-256 tumour-bearing (W). The rats were euthanised on three different gestational periods: at 12 days post-conception (dpc), at 16 dpc, and at 19 dpc. Serum, placenta and foetal metabolomic profiles were performed by ¹H-NMR spectra following the analyses using Chenomx NMR Analysis Software V8.3. The tumour evolution was exponential, affecting the placental metabolomic profile during all the pregnancy stages. The placental tissue in tumour-bearing dams developed at a lower speed, decreasing the foetus's weight. Associated with the serum metabolomic changes related to tumour growth, the placental metabolomic alterations impacted many metabolic pathways related to energy provision, protein synthesis and signalling, which directly harmed the foetus's development. The development of the foetus is clearly affected by the damage induced by the tumour evolution, which alters the metabolic profile of both the serum and the placenta, impairing early embryonic development.

Keywords: cancer during pregnancy; foetus; metabolism; placenta.

Figure 1

Placenta and foetus morphometric data from experimental groups, healthy (C) and tumour-bearing (W) dams at three distinct gestational periods, 12, 16, and 19 days post-conception: (a) placenta weight (g); (b) foetus weight (g); (c) ratio of foetal weight and placental weight. Data presented as mean ± standard deviation. Number of animals per group: C (placenta and foetus: 12 dpc n = 8; 16 dpc n = 8; 19 dpc n = 7); W group (placenta and foetus: 12 dpc n = 14; 16 dpc n = 12; 19 dpc n = 6). *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Figure 2

Maternal serum metabolite variations in healthy (C) and tumour-bearing (W) dams at three different gestational periods, 12-, 16- and 19-days post-conception. Number of animals per group: C at 12 dpc (n = 6), 16 dpc (n = 5) and 19 dpc (n = 6); W group at 12 dpc (n = 7), 16 dpc (n = 6) and 19 dpc (n = 6). For details see the Section 4. Legend: (a) 1-Methylhistidine; (b) Alanine; (c) Aspartate; (d) Betaine; (e) Carnitine; (f) Creatine; (g) Formate; (h) Fumarate; (i) Glutamine; (j) Glycine; (k) Lactate; (l) Leucine; (m) Phenylalanine; (n) Pyruvate; (o) Succinate; (p) Tryptophan; (q) Tyrosine; (r) Urea. The data were presented as the mean ± standard deviation. Statistical analysis was performed using a two-way ANOVA, and multiple comparisons were corrected using the post hoc test, specifically the multiple comparisons test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Figure 3

Placental metabolite variations in healthy (C) and tumour-bearing (W) dams at three different gestational periods, 12, 16 and 19 days post-conception. Number of animals per group: C at 12 dpc (n = 6), 16 dpc (n = 9) and 19 dpc (n = 6); W group at 12 dpc (n = 6),16 dpc (n = 8) and 19 dpc (n = 6). Legend: (a) 1-Methylhistidine; (b) Alanine; (c) Choline; (d) Creatine; (e) Fumarate; (f) Glucose; (g) Lactate; (h) Nicotinurate; (i) Pantothenate; (j) Pyruvate; (k) Serine; (l) Succinate; (m) Threonine; (n) Uridine; (o) Valine. The data were presented as the mean ± standard deviation. Statistical analysis was performed using a two-way ANOVA, and multiple comparisons were corrected using the post hoc test, specifically, the multiple comparisons test. Pregnancy: *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Figure 4

Foetal metabolite variations in healthy (C) and tumour-bearing (W) dams at three different gestational periods, 12, 16 and 19 days post-conception. Number of animals per group: C at 12 dpc (n = 6), 16 dpc (n = 6) and 19 dpc (n = 8); W group at 12 dpc (n = 8), 16 dpc (n = 7) and 19 dpc (n = 8). Legend: (a) Adenine; (b) Alanine; (c) AMP; (d) ADP; (e) ATP; (f) Choline; (g) Glucose; (h) Guanosine; (i) Glucose-6-phosphate; (j) Lactate; (k) IMP; (l) Succinate; (m) Nicotinurate; (n) UDP-Glucose. The data were presented as the mean ± standard deviation. Statistical analysis was performed using a two-way ANOVA, and multiple comparisons were corrected using the post hoc test, specifically the multiple comparisons test. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Figure 5

Experimental procedure to analyse serum, placental and foetal metabolomic profiles in pregnant tumour-bearing rats (W) compared to the healthy pregnant group (C). All the animals were euthanised at 12, 16 and 19 days of pregnancy. Illustration created using BioRender (accessed on 19 July 2023).