Identification of O-linked N-acetylglucosamine transferase (OGT) expression

in human placentas as a potential biomarker of prenatal stress exposure

Caroline Camilo1, Luana Martos Vieira1, Arleti Caramori Torrezan2, Antonia Beatriz Sousa1, Gisele Gouveia1, Veronica Euclydes1,
Aloísio Souza Felipe da Silva2,3, Alexandra Brentani4 ,Helena Brentani1
1 - Departamento & Instituto de Psiquiatria, Faculdade de Medicina FMUSP, Universidade de São Paulo, São Paulo, SP, BR; 2 - Hospital Universitário, Universidade de São Paulo, São Paulo, SP, BR;
3 - LIM/14, Faculdade de Medicina FMUSP, Universidade de São Paulo, São Paulo, SP, BR; 4 - Departamento de Pediatria, Faculdade de Medicina FMUSP, Universidade de São Paulo, São Paulo, SP, BR

BACKGROUND
O-linked N-acetylglucosamine transferase (OGT), also known as O-GlcNAc transferase, is an enzyme that catalyzes the O-GlcNAcylation post-translational modification in
nuclear and cytosolic proteins and plays an important role in metabolic, transcriptional and epigenetic processes. Animal model studies have proposed the placental OGT
expression as a promising biomarker of maternal stress exposure, identifying an association between lower levels of OGT in males, exposure to stress during pregnancy
and neurodevelopmental changes1. Here, we aimed to identify a differential OGT protein expression in human placenta tissues of males and females, using a birth
cohort characterized by prenatal exposure to environmental stress.
METHODS

Expression value 4 (E) - p: positivity;

ni: number of intensity categories;
Figure 1 – Representation of data collection and experiment steps. I: intensity of staining

RESULTS
A: Negative control
B: Weak intensity/positivity

Figure 4 – Linear regression model with OGT expression (E) and BW:PW ratio
between male and female placentas.
C: Moderate intensity/positivity

Figure 3 – Boxplots (T-Student Test) representing expression value (E) between

male and female placentas.

Table 1. Multivariate model with BMI-for-age as outcome

BMI-for-age
Predictors Estimates CI p-value
(Intercept) -8.89 -15.30 – -2.35 0.009
E OGT 0.02 0.00 – 0.03 0.018
D: Strong intensity/positivity

Sex [F] 10.50 4.33 – 16.67 0.001

BW:PW ratio -0.10 -0.30 – 0.09 0.283
Gestational age 0.05 -0.09 – 0.18 0.467
E OGT * Sex [F] -0.02 -0.04 - -0.01 0.002
R2 / R2 adjusted 0.297 / 0.209

Figure 2 – Detection of OGT protein expression in human placentas. Image illustrates the slides
visualization in the ImageScope 12.4.6 software and its analysis using the Pixel Count V9 algorithm
(Blue: Negative; Yellow: Weak intensity and positivity; Orange: Moderate intensity; Red: Strong
intensity). A: Negative control showing no staining; B,C and D: Positive staining in Figure 5 – Linear regression model with OGT expression (E) and BMI-for-age
syncytiotrophoblasts. Original magnification ×8 (Scale bar 300 μm). between male and female placentas.

DISCUSSION
Prenatal stress can affect placental metabolism and fetal growth, directly affecting the OGT regulatory pathway 5. We identified OGT expression in syncytiotrophoblasts,
the transporting epithelium of the human placenta which mediates maternal-fetal exchange of nutrients 6. No significant difference of OGT expression were found
between sex, but we found that BMI/age at birth was associated with OGT placenta expression in a sex specific manner. The lower was OGT expression in female
placentas, the higher was BMI/age. Our findings represent a progress in the hypothesis of OGT as a promising biomarker of male and female placentas response to
exposure to environmental risk factors. Further investigations using this approach in independent samples are necessary to replicate and explore the current findings.
REFERENCES
1 Howerton CL et al. Proc Natl Acad Sci U S A. 2013.;110(13):5169-74. 2 Brentani A et al. Int J Epidemiol. 2020;49(5):1438-1438g. 3 Dos Santos AC et al. BMC Public Health. 2021;21(1):865. 4 Silva AR et al. PLoS One. 2014;9(6):e99897
5 Hart B et al. J Mol Endocrinol. 2019;62(2):R155-R165. 6 Jansson T, Powell TL. Clin Obstet Gynecol. 2013;56(3):591-601.

Funding was provided by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP grants #2018/18560-6, #2020/14360-2 and #2021/00607-9)
and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq #310823/2021-8).

Conflict of interest: The authors declare that they have no conflict of interest. E-mail: carolcamilo@alumni.usp.br