Placental SARS-CoV-2 distribution correlates with level of tissue oxygenation in COVID-19-associated necrotizing histiocytic intervillositis/perivillous fibrin deposition.

INTRODUCTION: Recent evidence supports the - rare - occurrence of vertical transplacental SARS-CoV-2 transmission. We previously determined that placental expression of angiotensin-converting enzyme 2 (ACE2), the SARS-CoV-2 receptor, and associated viral cell entry regulators is upregulated by hypoxia. In the present study, we utilized a clinically relevant model of SARS-CoV-2-associated chronic histiocytic intervillositis/massive perivillous fibrin deposition (CHIV/MPFVD) to test the hypothesis that placental hypoxia may facilitate placental SARS-CoV-2 infection. METHODS: We performed a comparative immunohistochemical and/or RNAscope in-situ hybridization analysis of carbonic anhydrase IX (CAIX, hypoxia marker), ACE2 and SARS-CoV-2 expression in free-floating versus fibrin-encased chorionic villi in a 20-weeks' gestation placenta with SARS-CoV-2-associated CHIV/MPVFD. RESULTS: The levels of CAIX and ACE2 immunoreactivity were significantly higher in trophoblastic cells of fibrin-encased villi than in those of free-floating villi, consistent with hypoxia-induced ACE2 upregulation. SARS-CoV-2 showed a similar preferential localization to trophoblastic cells of fibrin-encased villi. DISCUSSION: The localization of SARS-CoV-2 to hypoxic, fibrin-encased villi in this placenta with CHIV/MPVFD suggests placental infection and, therefore, transplacental SARS-CoV-2 transmission may be promoted by hypoxic conditions, mediated by ACE2 and similar hypoxia-sensitive viral cell entry mechanisms. Understanding of a causative link between placental hypoxia and SARS-CoV-2 transmittability may potentially lead to the development of alternative strategies for prevention of intrauterine COVID-19 transmission.

Increased placental expression of angiotensin-converting enzyme 2, the receptor of SARS-CoV-2, associated with hypoxia in twin anemia-polycythemia sequence (TAPS).

INTRODUCTION: Recent reports suggest SARS-CoV-2, the virus causing COVID-19, may be transmittable from pregnant mother to placenta and fetus, albeit rarely. The efficacy of vertical transmission of SARS-CoV-2 critically depends on the availability of its receptor, ACE2, in the placenta. In the present study, we tested the hypothesis that placental ACE2 expression is oxygenation-dependent by studying the expression of ACE2 and associated cell entry regulators in the monochorionic twin anemia-polycythemia (TAPS) placenta, a model of discordant placental oxygenation. METHODS: We performed a retrospective comparative immunohistochemical, immunofluorescence and Western blot analysis of ACE2, TMPRSS2 and Cathepsin B expression in anemic and polycythemic territories of TAPS placentas (N = 14). RESULTS: ACE2 protein levels were significantly higher in the anemic twin territories than in the corresponding polycythemic territories, associated with upregulation of the key ACE2-related cell entry regulators, TMPRSS2 and Cathepsin B, immunolocalized to villous trophoblastic and stromal cells. Cellular colocalization of ACE2 and TMPRSS2, suggestive of functionality of this cell entry axis, was demonstrated by double immunofluorescence studies. DISCUSSION: Placental hypoxia is associated with upregulation of ACE2 expression, concomitant with increased expression of its key cell entry proteases. ACE2-regulated placental functions, both infection- and non-infection related, may be highly oxygenation-dependent.

Correlation between chorionic plate vascularization and risk of bronchopulmonary dysplasia in extremely preterm infants.

INTRODUCTION/OBJECTIVES: The chorionic plate vessels of the placenta are in direct continuity with the fetal vasculature, suggesting chorionic and fetal angiogenesis may be subjected to similar regulatory mechanisms. In this study, we determined the correlation between chorionic plate vascularization and complications of prematurity, focusing on bronchopulmonary dysplasia (BPD) and other conditions with important microvascular components. METHODS: We performed a clinicoplacental analysis of 127 extremely preterm infants (23-28 weeks gestation). Chorionic plate vascularization was assessed by number and density of perforating chorionic vessels (PCVs). Charts were reviewed for relevant maternal and neonatal data, including respiratory, neurologic and gastrointestinal complications of prematurity. RESULTS: The placentas displayed marked variability in number (36-523/placenta) and density of PCVs (0.46-3.74 PC V/cm2). The median PCV density of infants with severe BPD was significantly higher than that of infants without BPD (1.51 PC V/cm2 versus 1.09 PC V/cm2, P < 0.05). Conversely, the frequency of moderate-to-severe BPD was 33% higher in infants with PCV density ≥1.50 PC V/cm2 than in those with PCV density <1.50 PC V/cm2 (56% versus 40%, P < 0.01). There was no correlation with neonatal neurologic or gastrointestinal complications. CONCLUSION: Chorionic plate vascularization correlates with frequency and severity of BPD, supporting a vascular basis that in part is antenatal in origin. Quantitative assessment of chorionic plate vascularization may allow early identification of preterm infants at high risk for BPD (proposed threshold: PCV density ≥1.50 PC V/cm2). The lack of correlation between chorionic vascularization and neurologic/gastrointestinal complications suggests these conditions may have less important antenatal and/or vascular contributions.

Discordant placental oxygenation and autophagy in twin anemia-polycythemia sequence (TAPS).

BACKGROUND: (Macro)autophagy is an important process of self-degradation of macromolecules and organelles that ensures cellular homeostasis and energy preservation during stressful conditions. Dysregulated placental autophagy has been implicated in a wide range of pregnancy complications. Recent studies identified hypoxia as a key regulator of trophoblast autophagy in vitro; however, its effects on placental autophagy in vivo remain incompletely understood. In this study, we evaluated the monochorionic twin anemia-polycythemia sequence (TAPS) placenta as model of discordant placental oxygenation to determine the effects of hypoxia on placental autophagy in utero. METHODS: We performed a retrospective comparative analysis of tissue oxygenation and autophagy in anemic and polycythemic territories of TAPS placentas (N = 12). Archival tissues were subjected to immunohistochemical, immunofluorescence and Western blot analyses of carbonic anhydrase (CA) IX (hypoxia marker) and key autophagy/lysosomal markers. RESULTS: CAIX protein levels were significantly higher in anemic twin territories than in corresponding polycythemic territories, consistent with relative tissue hypoxia. Anemic placental shares further displayed significantly higher levels of LC3I/II (autophagosome markers) and LAMP1/2 (lysosome markers), associated with upregulated expression of lysosome/autophagosome activity-associated markers, transcription factor EB and cathepsin D. The accumulation of autophagosomes and lysosomes in anemic shares was accompanied by elevated p62 protein expression, suggestive of inhibition of the downstream autophagy pathway. CONCLUSIONS: TAPS placentas display striking intertwin discordance in tissue oxygenation and autophagic activity and may provide a suitable model for study of the interrelationship between hypoxia, autophagy, and pregnancy outcome in a monochorionic twin setting.

Redness discordance in monochorionic twin placentas: Correlation with clinical and placental findings.

INTRODUCTION/OBJECTIVES: Recent studies suggest redness (color) discordance of the placental basal plate may be a marker for twin anemia-polycythemia sequence (TAPS), a recently described complication of diamniotic-monochorionic twinning characterized by marked intertwin hemoglobin (Hb) discordance in the absence of oligohydramnios-polyhydramnios. In this study, we determined the clinicoplacental and choriovascular correlates of basal plate color discordance in monochorionic twin placentas, and assessed its value as postnatal indicator of TAPS. METHODS: We performed a clinicoplacental analysis of 100 consecutive non-TTTS diamniotic-monochorionic twin placentas with available photographic documentation of the basal plate. Basal plate redness was quantified by computer-assisted analysis of digital images and expressed as intertwin color difference ratio (CDR). RESULTS: The CDR ranged between 1.00 and 3.58 (median CDR: 1.14; 90th %ile: 1.98). Compared to twins with low CDR (N = 90), twins with high CDR (≥2.0; N = 10) had significantly higher hemoglobin difference (11.25 g/dL versus 2.55 g/dL) and significantly fewer and smaller artery-to-artery (AA) and artery-to-vein (AV) anastomoses. Apgar scores and birth weights were equivalent in both groups. Among the 10 twin sets with high CDR, six (60%) qualified as TAPS, as defined by intertwin Hb difference >8 g/dL and absent or very small AA and AV anastomoses. Conversely, 6 of 8 (75%) twin sets with TAPS had a CDR ≥ 2.0. CONCLUSION: Intertwin CDR correlates with intertwin hemoglobin difference and chorionic angioarchitecture. A CDR value ≥ 2.0 (the 90%ile value for CDR derived from the present cohort) has high specificity (96%), but relatively low positive predictive value (60%) as indicator of TAPS, as currently defined.

Florid Intussusceptive-like Microvascular Dysangiogenesis in a Preterm Lung.

The cellular mechanisms underlying the microvascular dysangiogenesis of bronchopulmonary dysplasia (chronic lung disease of the newborn) remain largely undetermined. We report unusual pulmonary vascular findings in a 27-week-gestation male newborn who died on the second day of life from intractable respiratory failure, following a pregnancy complicated by prolonged membrane rupture and persistent severe oligohydramnios. As expected, postmortem examination revealed pulmonary hypoplasia (lung/body weight ratio: 2.23%; 10th percentile for 27 weeks: 2.59%). In addition, lung microscopy revealed complex networks of non-sprouting, tortuous, and bulbously dilated capillaries, randomly distributed in widened airspace septa. Anti-smooth muscle actin immunohistochemistry demonstrated immunoreactive central densities within capillary lumina, suggestive of intravascular pillar formation. The plexus-forming, non-sprouting type of angiogenesis and apparent transluminal pillar formation are consistent with intussusceptive ("longitudinal splitting") angiogenesis. In concordance with previous observations made in human fetal lung xenografts, these findings support the notion that human postcanalicular lungs have the capacity to switch from sprouting to non-sprouting, intussusceptive-like angiogenesis, possibly representing an adaptive response activated by hemodynamic flow alterations and/or hypoxia. The possible relationship between the intussusceptive-like vascular changes observed in this case and the microvascular dysangiogenesis characteristic of bronchopulmonary dysplasia remains to be determined.

Postmortem lung volume/body weight standards for term and preterm infants.

Assessment of lung growth is a critical component of the perinatal autopsy. Increased lung liquid content may lead to overestimation of lung growth based on (wet) lung weight. In contrast, lung volume is not influenced by intraalveolar lung liquid. Our aim was to establish age-specific reference values for postmortem lung volume/BW in preterm and term infants. We performed a retrospective analysis of fetuses/infants (16-41 weeks' gestation) without (N = 134) or with (N = 79) risk factors for pulmonary hypoplasia. Lungs were inflated at standardized pressure and volumes determined by water immersion method. Lung volume increased 11-fold between 16 and 41 weeks' gestation, concomitant with a 16-fold increase in BW. Mean lung volume/BW remained constant at 33-34 ml/kg between 16 and 31 weeks' gestation and decreased to 23.4 ml/kg at term. Lung volume/BW of infants with severe risk factors (renal anomalies, diaphragmatic hernia) was significantly lower than age-matched standards. In this group, all fetuses/infants diagnosed as having lung hypoplasia by lung volume/BW also had lung hypoplasia LW/BW standards. However, in infants with "softer" risk factors (rupture of membranes, chromosomal anomalies), 5/26 cases diagnosed with lung hypoplasia based on lung volume/BW had normal LW/BW ratios. In these discrepant cases, lung sections showed significant inflammation and edema, likely accounting for increased wet lung weight. In conclusion, we determined age-specific lung volume/BW reference values for preterm and term infants. In selected situations assessment of lung volume/BW may represent a useful complementary tool to LW/BW for postmortem evaluation of lung size.

Correlation between cord insertion type and chorionic villus vascularization of the co-twin in diamniotic-monochorionic twin pregnancies.

BACKGROUND: Recent evidence suggests that cord insertion type of one twin correlates with chorionic plate vascularization of the monochorionic co-twin. Specifically, for twins with paracentral cords, chorionic plate vascularization is significantly greater when the co-twin has a velamentous, rather than paracentral cord insertion. AIMS: To determine whether this correlation between cord insertion type and vascularization of the co-twin also extends to the deeper chorionic villus tree. STUDY DESIGN: Morphometric analysis of chorionic villus vascularization in CD31-immunostained sections of a retrospective cohort of gestational age-matched third trimester monochorionic placentas with discordant paracentral/velamentous (PC/V) or concordant paracentral/paracentral (PC/PC) cord insertions. OUTCOME MEASURES: Vascular numerical density (number of vascular profiles per unit villus stromal area) of intermediate villi (>80 µm diameter) and terminal villi (<80 µm). RESULTS: For twins with paracentral cord insertion, the vascular numerical density of intermediate villi was significantly higher for twins in a discordant PC/V relationship than for those in a concordant PC/PC relationship (P<0.05), thus replicating previous findings in superficial chorionic vessels. For terminal villi, in contrast, the vascular numerical density of twins with paracentral cords in a PC/V combination was significantly lower than of those in a PC/PC combination, and similar to that of their co-twins with velamentous cord insertion. CONCLUSIONS: Early placental angiogenesis in monochorionic twin gestations may be influenced by implantation and cord localization of the co-twin. The regulation of terminal villus angiogenesis appears to be dissociated from more proximal villus angiogenesis and independent of cord insertion of the co-twin.

Intrauterine demise due to congenital mesoblastic nephroma in a fetus conceived by assisted reproductive technology.

OBJECTIVE: To report a case of antenatally diagnosed congenital mesoblastic nephroma in an assisted reproductive technology (ART) conception. DESIGN: Case report. SETTING: Tertiary care university-affiliated hospital. PATIENT(S): Fetus of 26-weeks' gestation with antenatally diagnosed large abdominal tumor. INTERVENTION(S): ART with transfer of cryopreserved embryo. MAIN OUTCOME MEASURE(S): Postmortem examination. RESULT(S): Examination revealed a congenital mesoblastic nephroma, mixed classic and cellular type, with marked intratumoral hemorrhage and associated hydrops. The marked fetal erythroblastosis was suggestive of fetal response to pronounced anemia. Intrauterine demise is attributed to fetal intratumoral hemorrhage and early nonimmune hydrops secondary to a large congenital mesoblastic nephroma. CONCLUSION(S): This is the third reported case of congenital mesoblastic nephroma in an ART conception. Whether the association between mesoblastic nephroma and ART is coincidental or causative remains to be determined.