Sex-specific differences in the severity of pulmonary hypoplasia in experimental congenital diaphragmatic hernia and implications for extracellular vesicle-based therapy

Fabian Doktor; Emily Lo; Victoria Fortuna; Kasra Khalaj; Miguel Garcia; Rebeca Lopes Figueira; Martin Lacher; Lina Antounians; Augusto Zani

doi:10.1007/s00383-024-05856-0

Sex-specific differences in the severity of pulmonary hypoplasia in experimental congenital diaphragmatic hernia and implications for extracellular vesicle-based therapy

Pediatr Surg Int. 2024 Oct 28;40(1):278. doi: 10.1007/s00383-024-05856-0.

Authors

Fabian Doktor^{1

2

3}, Emily Lo^{1

2

4}, Victoria Fortuna^{1

2}, Kasra Khalaj^{1

2}, Miguel Garcia^{1

2}, Rebeca Lopes Figueira^{1

2}, Martin Lacher³, Lina Antounians^{1

2}, Augusto Zani^{5

6

7}

Affiliations

¹ Developmental and Stem Cell Biology Program, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children, Toronto, M5G 0A4, Canada.
² Division of General and Thoracic Surgery, The Hospital for Sick Children, Toronto, M5G 1X8, Canada.
³ Department of Pediatric Surgery, University of Leipzig, 04109, Leipzig, Germany.
⁴ Division of Neonatology, Department of Paediatrics, The Hospital for Sick Children, University of Toronto, Toronto, M5G 1X8, Canada.
⁵ Developmental and Stem Cell Biology Program, Peter Gilgan Centre for Research and Learning, The Hospital for Sick Children, Toronto, M5G 0A4, Canada. [email protected].
⁶ Division of General and Thoracic Surgery, The Hospital for Sick Children, Toronto, M5G 1X8, Canada. [email protected].
⁷ Department of Surgery, University of Toronto, Toronto, M5T 1P5, Canada. [email protected].

PMID: 39467854
DOI: 10.1007/s00383-024-05856-0

Abstract

Purpose: Amniotic fluid stem cell extracellular vesicles (AFSC-EVs) hold regenerative potential to treat hypoplastic lungs secondary to congenital diaphragmatic hernia (CDH). This study aims to investigate sex-specific differences in pulmonary hypoplasia severity and responses to AFSC-EV administration in an experimental CDH mouse model.

Methods: C57BL/6J dams were fed with nitrofen + bisdiamine (left-sided CDH) or olive oil only (control) at embryonic day (E) 8.5. Lungs were dissected (E18.5), grown ex vivo and treated with medium ± AFSC-EVs that were collected via ultracentrifugation and characterized (nanoparticle tracking analysis, electron microscopy, Western blotting). Pulmonary hypoplasia was assessed via mean linear intercept (MLI). Gene and protein expression changes (Cd31, Enos, Il1b, TNFa) were measured via RT-qPCR and immunofluorescence. Pups were genotyped for Sry.

Results: Experimental CDH showed a male predominance without sex differences for pulmonary hypoplasia severity, fetal lung vascularization, and inflammation. AFSC-EV administration led to improved lung growth (decreased MLI), improved fetal lung vascularization (increased Cd31 and Enos), and decreased fetal lung inflammation (Il1b, TNFa). There was no sex-specific response to AFSC-EV administration.

Conclusion: This study shows sex-independent impaired lung growth, vascularization and fetal lung inflammation in a CDH mouse model. Antenatal administration of AFSC-EVs reverses aspects of pulmonary hypoplasia secondary to CDH independent of the biological sex.

Keywords: Exosomes; Fetal medicine; Lung inflammation; Pulmonary hypoplasia; Sex differences; Stem cells.

MeSH terms

Abnormalities, Multiple
Amniotic Fluid
Animals
Disease Models, Animal*
Extracellular Vesicles* / metabolism
Female
Hernias, Diaphragmatic, Congenital* / complications
Hernias, Diaphragmatic, Congenital* / genetics
Lung Diseases
Lung* / abnormalities
Lung* / embryology
Male
Mice
Mice, Inbred C57BL*
Pregnancy
Respiratory System Abnormalities / genetics
Severity of Illness Index
Sex Factors

Supplementary concepts

Lung agenesis

Abstract

MeSH terms

Supplementary concepts

Grants and funding