One of the biggest challenges for babies with CDH is that fetal hypoplastic lungs are immature, whereby there is an impairment of cell differentiation, especially in the epithelium.
And so moving forward, these results have direct clinical relevance as an AFSEV treatment represents a promising self-free approach to improving lung growth in babies with CDH.
Many refer to autophagy as a recycling mechanism of cellular tra, so to speak, and this is because it's one of the key ways that cell survival is promoted.
At the protein level in human tissue, all three autophagy markers (Beclin 1, ATG5, sequestosome) were dysregulated in the hypoplastic group, and human AFSCEVs restored all three markers back to control
CAPS - Administration of amniotic fluid stem cell extracellular vesicles regenerates the lung epithelium in fetal rats with CDH at translationally relevant developmental stages - Kasra Khalaj
▶Ep 12 · 0:29
quoteCongenital diaphragmatic hernia is characterized by impaired fetal lung growth and maturation. This condition has unacceptably high mortality and morbidity rates.↗
▶Ep 12 · 0:29
clinicalCongenital diaphragmatic hernia is characterized by impaired fetal lung growth and maturation with unacceptably high mortality and morbidity rates.↗
▶Ep 12 · 1:00
quoteOne of the biggest challenges for babies with CDH is that fetal hypoplastic lungs are immature, whereby there is an impairment of cell differentiation, especially in the epithelium.↗
▶Ep 12 · 1:00
clinicalOne of the biggest challenges for babies with CDH is that fetal hypoplastic lungs are immature with impairment of cell differentiation, especially in the epithelium.↗
▶Ep 12 · 1:15
clinicalExtracellular vesicles (EVs) are the main mediators of stem cell paracrine signaling.↗
▶Ep 12 · 1:25
clinicalIn a recently published proof of concept study, amniotic fluid stem cell derived EVs can rescue fetal lung growth and maturation in several models of CDH at the pseudoglandular stage, which corresponds to 7 to 16 weeks of human gestation.↗
▶Ep 12 · 1:55
clinicalCDH is diagnosed around 18 to 20 weeks of human gestation.↗
▶Ep 12 · 1:55
clinicalThe canalicular and saccular stages are translationally relevant developmental stages for CDH intervention, corresponding to the time when diagnosis occurs.↗
▶Ep 12 · 2:15
clinicalThe nitrofen model is a well-established model for studying CDH, created by administering the herbicide nitrofen to pregnant rat dams at embryonic day 9.5.↗
▶Ep 12 · 2:35
clinicalKey epithelial populations in the fetal lung include alveolar type 1 and type 2 cells (primarily comprising the alveolus) and club, ciliated, and basal cells (mainly comprising the bronchi and bronchioles).↗
▶Ep 12 · 2:55
clinicalHypoplastic lungs at canalicular and saccular stages show downregulation of alveolar type 1 and 2 cells, as well as basal and club cells compared to control lungs.↗
▶Ep 12 · 3:10
clinicalTreatment of hypoplastic lungs with amniotic fluid stem cell derived extracellular vesicles restores primary epithelial cell markers, indicating that cell homeostasis can be achieved.↗
▶Ep 12 · 4:42
clinicalIn vivo experiments using intraamniotic and tracheal administration routes of extracellular vesicles in rat models have shown promising results.↗
▶Ep 12 · 5:36
clinicalSafety and feasibility studies of AFSC-EV treatment are being investigated in collaboration with Jan Deprest in a lamb model.↗
▶Ep 12 · 6:33
clinicalCiliated cells in hypoplastic CDH lungs did not show rescue with AFSC-EV treatment, which may reflect achievement of a different homeostatic cell composition rather than failure of the therapy.↗
▶Ep 12 · 7:00
clinicalKey markers of branching morphogenesis are being rescued effectively by AFSC-EV treatment.↗
EUPSA - MicroRNAs in amniotic fluid stem cellextracellular vesicles modulate lung development in experimental congenital diaphragmatic hernia - Kasra Khalaj
▶Ep 13 · 0:31
clinicalHypoplastic lungs of babies with pulmonary hypoplasia secondary to CDH have impaired fetal lung growth and maturation↗
▶Ep 13 · 0:31
quoteIt is well known that hypoplastic lungs of babies with pulmonary hypoplasia secondary to CDH have impaired fetal lung growth and maturation.↗
▶Ep 13 · 0:39
clinicalSeveral treatment agents have been administered prenatally for CDH, but none have been shown to fully rescue lung development↗
▶Ep 13 · 0:45
clinicalExtracellular vesicles are nanoparticles known to be the mediators of stem cell paracrine signaling↗
▶Ep 13 · 0:56
clinicalAFSCEV administration rescued the number of lung branches back to normal in CDH models↗
▶Ep 13 · 1:07
clinicalAFSCEVs improve fetal lung maturation as shown by rescuing the expression of surfactant protein C↗
▶Ep 13 · 1:17
clinicalAFSCEVs contain microRNA 17-92 cluster, which has been reported in the literature to modulate fetal lung development↗
▶Ep 13 · 1:31
clinicalKnockout of the microRNA 17-92 cluster can recapitulate pulmonary hypoplasia↗
▶Ep 13 · 1:46
clinicalAutophagy has been shown to be critical for lung branching morphogenesis↗
▶Ep 13 · 1:55
clinicalAutophagy is a recycling mechanism of cellular trash and is one of the key ways that cell survival is promoted↗
▶Ep 13 · 1:55
quoteMany refer to autophagy as a recycling mechanism of cellular tra, so to speak, and this is because it's one of the key ways that cell survival is promoted.↗
▶Ep 13 · 2:04
clinicalAutophagy is most impaired at the pseudoglandular and canalicular stages in the nitrofen model of CDH↗
▶Ep 13 · 2:18
clinicalHigh levels of sequestosome indicate autophagy impairment↗
▶Ep 13 · 2:18
clinicalCDH fetal lungs show reduced levels of Beclin 1 and ATG5 (autophagy activators) and higher levels of sequestosome (autophagy adapter) at pseudoglandular and canalicular stages↗
▶Ep 13 · 2:32
clinicalTreatment with AFSCEVs restores autophagy key genes (Beclin 1, ATG5, sequestosome) at the pseudoglandular and canalicular stage↗
▶Ep 13 · 2:56
clinicalAFSCEVs are taken up by virtually all cells in fetal lung explants, including the fetal lung epithelium↗
▶Ep 13 · 3:19
clinicalActivation of autophagy by AFSCEVs is localized in the lung epithelial compartment of the fetal lung microenvironment↗
▶Ep 13 · 3:27
clinicalKnockdown of microRNAs 17 and 20 in AFSCEVs results in increased sequestosome and down regulation of Beclin mRNA, indicating impaired autophagy↗
▶Ep 13 · 3:53
clinicalKnockdown of microRNAs 17 and 20 results in lower levels of LC3B2 protein, which is the active isoform of the autophagy cascade↗
▶Ep 13 · 4:03
quoteWell, by now I bet you're all asking, well, are these findings clinically relevant in human CDH?↗
▶Ep 13 · 4:10
clinicalHuman fetal lung explant model of pulmonary hypoplasia was established using fetal lungs from healthy terminated fetuses from 15 to 19 weeks of gestation, corresponding to late pseudoglandular and canalicular stages↗
▶Ep 13 · 4:37
clinicalIn human hypoplastic fetal lung explants, there is a reduction of Beclin 1 and ATG5 at the mRNA level↗
▶Ep 13 · 4:58
clinicalTreatment with human AFSCEVs results in restoration of Beclin 1 and ATG5 in human hypoplastic fetal lung explants↗
▶Ep 13 · 5:04
clinicalAt the protein level in human tissue, all three autophagy markers (Beclin 1, ATG5, sequestosome) were dysregulated in the hypoplastic group, and human AFSCEVs restored all three markers back to control↗
▶Ep 13 · 5:17
opinionThis is the first study discovering autophagy impairment as an important mechanism in CDH pathophysiology↗
▶Ep 13 · 5:17
quoteIn conclusion, we are very excited as this is the first study discovering an important mechanism in the CDH pathophysiology.↗
▶Ep 13 · 5:24
clinicalAutophagy levels are restored with administration of AFSCEVs, thus partially explaining their effect on branching morphogenesis↗
▶Ep 13 · 5:31
clinicalAutophagy can be targeted with microRNAs 17 and 20↗
▶Ep 13 · 7:39
opinionFuture work includes looking at ER stress, which is very closely interrelated with autophagy pathway, to see if AFSCEVs can exert an effect on this related mechanism↗
▶Ep 13 · 8:07
clinicalStudies in oncology have shown that the quantity of extracellular vesicles released when autophagy is impaired is different↗
▶Ep 13 · 8:37
opinionFuture functional studies will examine endogenous extracellular vesicle production in hypoplastic fetal lungs↗
Kasra Khalaj, MSc, PhD - Best of the Best in Pediatric Surgery 2024
▶Ep 30 · 0:22
quoteIt is well known that the lungs of babies with pulmonary hyperplasia, secondary to congenital diaphragmatic hernia, have the impaired triad of factors with impaired fetal lung growth, maturation, and vascularization.↗
▶Ep 30 · 0:22
clinicalLungs of babies with pulmonary hypoplasia secondary to congenital diaphragmatic hernia have an impaired triad of factors: impaired fetal lung growth, maturation, and vascularization.↗
▶Ep 30 · 1:00
clinicalPrenatal therapies administered to treat pulmonary hypoplasia have not been able to fully rescue lung development.↗
▶Ep 30 · 1:00
quoteAlthough some therapies have been administered prenatally to treat pulmonary hyperplasia, none of these have been able to fully rescue lung development.↗
▶Ep 30 · 1:20
clinicalExtracellular vesicles are particles responsible for paracrine signaling.↗
▶Ep 30 · 1:40
quoteI'm excited to show you all the novel human fetal lung eggplant model which we've now established at Sickkis.↗
▶Ep 30 · 2:20
clinicalRAC1 is a Rho GTPase that modulates lung branching morphogenesis.↗
▶Ep 30 · 2:40
clinicalRAC1 expression is down-regulated in CDH autopsy specimens compared to age-matched controls.↗
▶Ep 30 · 3:20
clinicalFetal lungs treated with RAC1 inhibitor display histological features resembling those of pulmonary hypoplasia.↗
▶Ep 30 · 3:40
clinicalHuman AFSCEVs restore airspace density with decreased parenchyma in hypoplastic fetal lungs.↗
clinicalHuman AFSCEVs restore key markers responsible for fetal lung maturation: surfactant protein C and podoplanin.↗
▶Ep 30 · 4:20
clinicalHuman AFSCEV-treated explants had reduced medial wall thickness and restored vascular density.↗
▶Ep 30 · 4:45
clinicalA unique epithelial cell cluster termed 'stressed AT1/AT2' had high representation in hypoplastic lungs and was down-regulated after AFSCEV administration.↗
▶Ep 30 · 5:00
clinicalThe stressed AT1/AT2 cluster had up-regulated signaling pathways involved in cellular stress, death, and inflammation.↗
▶Ep 30 · 5:10
clinicalA cluster termed 'developing AT1/AT2 cells' had the highest proportion in hypoplastic lungs treated with AFSCEVs and showed high TGF-beta and VEGF signaling.↗
▶Ep 30 · 5:25
clinicalHuman AFSCEV administration reduces the number of myofibroblasts in hypoplastic fetal lungs.↗
▶Ep 30 · 5:35
clinicalHuman AFSCEV administration restores vascular endothelial cluster nuclei numbers back to control levels.↗
▶Ep 30 · 5:50
clinicalThe restored vascular endothelial cluster had increased VEGF signaling, which is a primary pathway involved in angiogenesis.↗
▶Ep 30 · 6:05
clinicalThis is the first study to describe that human AFSCEV administration promotes the triad of fetal lung growth, maturation, and vascularization in fetal hypoplastic lungs.↗
▶Ep 30 · 6:05
quoteSo in conclusion, we are very excited as it is the first study to describe that human AFSCEV administration promotes the triad of factors, fetal lung growth, maturation, and vascularization in fetal hypoplastic lungs.↗
▶Ep 30 · 6:20
quoteAnd so moving forward, these results have direct clinical relevance as an AFSEV treatment represents a promising self-free approach to improving lung growth in babies with CDH.↗
▶Ep 30 · 6:35
clinicalInvestigators have tested various routes of AFSCEV administration including intratracheal and maternal IV in animal models.↗
▶Ep 30 · 6:50
clinicalIntra-amniotic injection has shown promising results for AFSCEV administration.↗
▶Ep 30 · 7:00
clinicalIn the lamb CDH model, AFSCEVs are being administered in combination with fetal endoscopic tracheal occlusion (FETO).↗
Kasra's statements about Congenital Diaphragmatic Hernia31 statements
EUPSA - MicroRNAs in amniotic fluid stem cellextracellular vesicles modulate lung development in experimental congenital diaphragmatic hernia - Kasra Khalaj
▶Ep 11 · 0:31
quoteIt is well known that hypoplastic lungs of babies with pulmonary hypoplasia secondary to CDH have impaired fetal lung growth and maturation.↗
▶Ep 11 · 0:31
clinicalHypoplastic lungs of babies with pulmonary hypoplasia secondary to CDH have impaired fetal lung growth and maturation↗
▶Ep 11 · 0:39
clinicalSeveral treatment agents have been administered prenatally for CDH, but none have been shown to fully rescue lung development↗
▶Ep 11 · 0:45
clinicalExtracellular vesicles are nanoparticles known to be the mediators of stem cell paracrine signaling↗
▶Ep 11 · 0:56
clinicalAFSCEV administration rescued the number of lung branches back to normal in CDH models↗
▶Ep 11 · 1:07
clinicalAFSCEVs improve fetal lung maturation as shown by rescuing the expression of surfactant protein C↗
▶Ep 11 · 1:17
clinicalAFSCEVs contain microRNA 17-92 cluster, which has been reported in the literature to modulate fetal lung development↗
▶Ep 11 · 1:31
clinicalKnockout of the microRNA 17-92 cluster can recapitulate pulmonary hypoplasia↗
▶Ep 11 · 1:46
clinicalAutophagy has been shown to be critical for lung branching morphogenesis↗
▶Ep 11 · 1:55
quoteMany refer to autophagy as a recycling mechanism of cellular tra, so to speak, and this is because it's one of the key ways that cell survival is promoted.↗
▶Ep 11 · 1:55
clinicalAutophagy is a recycling mechanism of cellular trash and is one of the key ways that cell survival is promoted↗
▶Ep 11 · 2:04
clinicalAutophagy is most impaired at the pseudoglandular and canalicular stages in the nitrofen model of CDH↗
▶Ep 11 · 2:18
clinicalCDH fetal lungs show reduced levels of Beclin 1 and ATG5 (autophagy activators) and higher levels of sequestosome (autophagy adapter) at pseudoglandular and canalicular stages↗
▶Ep 11 · 2:18
clinicalHigh levels of sequestosome indicate autophagy impairment↗
▶Ep 11 · 2:32
clinicalTreatment with AFSCEVs restores autophagy key genes (Beclin 1, ATG5, sequestosome) at the pseudoglandular and canalicular stage↗
▶Ep 11 · 2:56
clinicalAFSCEVs are taken up by virtually all cells in fetal lung explants, including the fetal lung epithelium↗
▶Ep 11 · 3:19
clinicalActivation of autophagy by AFSCEVs is localized in the lung epithelial compartment of the fetal lung microenvironment↗
▶Ep 11 · 3:27
clinicalKnockdown of microRNAs 17 and 20 in AFSCEVs results in increased sequestosome and down regulation of Beclin mRNA, indicating impaired autophagy↗
▶Ep 11 · 3:53
clinicalKnockdown of microRNAs 17 and 20 results in lower levels of LC3B2 protein, which is the active isoform of the autophagy cascade↗
▶Ep 11 · 4:03
quoteWell, by now I bet you're all asking, well, are these findings clinically relevant in human CDH?↗
▶Ep 11 · 4:10
clinicalHuman fetal lung explant model of pulmonary hypoplasia was established using fetal lungs from healthy terminated fetuses from 15 to 19 weeks of gestation, corresponding to late pseudoglandular and canalicular stages↗
▶Ep 11 · 4:37
clinicalIn human hypoplastic fetal lung explants, there is a reduction of Beclin 1 and ATG5 at the mRNA level↗
▶Ep 11 · 4:58
clinicalTreatment with human AFSCEVs results in restoration of Beclin 1 and ATG5 in human hypoplastic fetal lung explants↗
▶Ep 11 · 5:04
clinicalAt the protein level in human tissue, all three autophagy markers (Beclin 1, ATG5, sequestosome) were dysregulated in the hypoplastic group, and human AFSCEVs restored all three markers back to control↗
▶Ep 11 · 5:17
quoteIn conclusion, we are very excited as this is the first study discovering an important mechanism in the CDH pathophysiology.↗
▶Ep 11 · 5:17
opinionThis is the first study discovering autophagy impairment as an important mechanism in CDH pathophysiology↗
▶Ep 11 · 5:24
clinicalAutophagy levels are restored with administration of AFSCEVs, thus partially explaining their effect on branching morphogenesis↗
▶Ep 11 · 5:31
clinicalAutophagy can be targeted with microRNAs 17 and 20↗
▶Ep 11 · 7:39
opinionFuture work includes looking at ER stress, which is very closely interrelated with autophagy pathway, to see if AFSCEVs can exert an effect on this related mechanism↗
▶Ep 11 · 8:07
clinicalStudies in oncology have shown that the quantity of extracellular vesicles released when autophagy is impaired is different↗
▶Ep 11 · 8:37
opinionFuture functional studies will examine endogenous extracellular vesicle production in hypoplastic fetal lungs↗
