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Endometrial perivascular cells establish unique cytokine gradients in decidual process
A10
Abstracts / Placenta 35 (2014) A1eA23
O-026. PLACENTAL AMINO ACID TRANSPORTER FUNCTION ADAPTS TO MATERNAL AND FETAL NUTRIENT CONDITIONS Eiji Shibata David, J. Askew, Yukiyo Aiko, Chiharu Tomonaga, Satoshi Aramaki, Hirohide Inagaki, Toru Hachisuga. Department of Obstetrics and Gynecology, University of Occupational and Environmental Health, Japan Background: Studies suggest that placental nutrient uptake activity is responsive to both maternal and fetal nutrient demands. We hypothesize that under conditions of limited nutrient availability to the fetus, as often present in preeclampsia (PE), IUGR, and insufficient weight-gain during pregnancy, a general adaptive response aimed to increase amino acid transport activity may be observed in the placenta. Method: A total of 40 placentas from full-term (n¼10) and pre-term (n¼10) normal pregnancies, IUGR (n¼10), and PE (n¼10) associated pregnancies were looked at by immunohistochemistry followed by relative qualitative scoring to compare expression levels and localization of System L, ASCT2, and mTOR proteins. Result: Syncytiotrophoblast (ST) in placenta of pregnancies complicated by IUGR or PE showed significant increases in the levels of 4F2hc and LAT1 compared to both full-term control and pre-term pregnancies seperately. Elevated mTOR protein was uniquely higher in IUGR placentas compared to full-term controls. Total cellular ASCT2 transporter protein levels were similar in all groups, however, levels of ASCT2 protein localized to the ST microvillous membrane (MVM) were significantly lower in IUGR compared to both full-term and pre-term pregnancies. Additionally, ASCT2 and mTOR protein levels were positively associated with maternal prepregnancy BMI. Conclusion: First, in conditions of limited nutrient availability, such as PE or IUGR, there is an overall increase in the level of System L and mTOR protein expression in the ST, suggestive of an adaptive response. Second, a decrease in ASCT2 protein at the ST MVM suggests a post-translational event that may decrease AAT activity in IUGR placentas. Third, a physiological link between transporter expression and pre-pregnancy BMI is suggested based upon a positive association observed with ASCT2 and mTOR expression values.
O-027. ENDOMETRIAL PERIVASCULAR CELLS ESTABLISH UNIQUE CYTOKINE GRADIENTS IN DECIDUAL PROCESS a
a
a
Keisuke Murakami , Keiji Kuroda , Shintaro Makino , Atsuo Itakura a, Satoru Takeda a, Jan J. Brosens b. a Department of Obstetrics and Gynaecology, Juntendo University Faculty of Medicine, Japan; b Division of Reproductive Health, Clinical Science Research Laboratories, Warwick Medical School, University of Warwick, Japan Aim: The endometrial perivascular microenvironment is rich in mesenchymal stem-like cells that express type 1 integral membrane protein Sushi domain containing 2 (SUSD2) but the role of these cells in the decidual transformation of this tissue in pregnancy is unknown. We used an antibody directed against SUSD2 to isolate perivascular (SUSD2+) and non-perivascular (SUSD2-) fibroblasts from mid-luteal biopsies. Method: SUSD2+ and SUSD2- cells were purified from mid-luteal endometrial biopsies by magnetic bead separation and each cell population were expanded in vitro, then, subjected to RNA-seq analysis or differentiation experiments induced by cAMP and MPA to assess secretome profile. Result: On average, positive cells comprised 6.8% of endometrial stromal cells and 6-fold enriched in clonogenic cells. RNA sequencing revealed that cultures derived from SUSD2+ progenitor cells remain phenotypically distinct by the enrichment of novel and established endometrial perivascular signature genes including AOC3, NOTCH3, ELN, MYH11, DES, ACTA2, MCAM, and CSPG4. Decidualization transformed perivascular SUSD2+ -derived cells into the dominant source of a range of chemokines and cytokines, including CCL7, LIF and LTA.
Conclusion: Our findings confirmed that SUSD2+ -derived cells remain the expression of perivascular signature genes even after in-vitro culture. Decidual response is spatially organized at the feto-maternal interface with differentiating perivascular cells establishing distinct cytokine and chemokine gradients that could have the key role in decidual process.
O-028. LOCALIZATION OF TRPV5 VANILLOID RECEPTOR 5) TROPHOBLASTS
(TRANSIENT RECEPTOR PROTEIN IN HUMAN
POTENTIAL PLACENTAL
Yoko Aoyama, David J. Askew, Yukiyo Aiko, Chiharu Tomonaga, Satoshi Aramaki, Hirohide Inagaki, Toru Hachisuga, Eiji Shibata. Department of Obstetrics and Gynecology, University of Occupational and Environmental Health, Japan The Transient Receptor Proteins TRPV 5 and TRPV6 are members of a highly conserved family of Calcium channels sensitive to temperature, proteases, chemicals, shear stress, as well as intracellular Ca2+. TRPV 5 and 6 are both highly selective for Ca2+ as well as certain metal ions including Cadmium and Zinc. TRPV 5 and 6 are expressed in the placenta, and are thought to be the primary Ca2+ channels of the syncytiotroplast, and responsible for active Ca2+ transport across the placenta. Ca2+ transport is critical for proper fetal development, particularly bone calcification. Recent reports describe reduced activity and expression of these Ca2+ channels in the trophoblast of Preeclampsia-associated placenta. However, only TRPV6 has been shown to be expressed in the syncytiotrophoblast. Using immunohistochemistry, we found that TRPV 5 protein expression is primarily expressed in the cytotrophoblasts of the microvillous. TRPV5 staining in the syncytiotrophblast was very limited, primarily restricted to the nuclear syncytium. Therefore, the localization of TRPV5 with the placental microvillous is quite different from TRPV 6. In addition, our preliminary studies found no significant differences in TRPV 5 expression levels in the placenta of Preeclampsia or IUGR- associated pregnancies compared to controls. Our data suggests that TRPV5 and TRPV6 have distinct roles in the placenta.
O-029. THE NEW APPROACH TO DIAGNOSIS AND EVALUATE OF PLACENTA ACCRETE USING 3D SLICER Rie Oyama, Chizuko Isurugi, Shino Tanaka, Tomoyuki Fukagawa, Ikue Nakayama, Yuri Sasaki, Tomonobu Kanasugi, Aikihiko Kikuchi, Toru Sugiyama. Iwate Medical University Department Obstetrics and Gynecology, Japan Introduction: placenta accrete is the most important disease of the obstetric filed. In particular, in the case of a placenta that is attached over the side to posterior uterine in third trimester, it is necessary to evaluate the location of placenta sites, Therefore, we use various applications of ultrasound equipment, and combination with MR image. So, we tried to evaluate the identification of the attachment site of placenta accrete using 3D Slicer. 3D Slicer is open source -medical image software, which based at Harvard Medical School that consists of more than over 370k lines of code, mostly C++. This massive software development effort has been enabled by the participation of several large scale. NIH funded efforts, including the NA-MIC, NAC, BIRN, CIMIT and NCIGT communities (www.slicer.org). This software could able to evaluate the region of interest in the placenta attached uterine wall, and to create reconstructing image (for example: using affine transformation). 3D reconstructing image have processed on DICOM data (Raw volume data) of ultrasound, CT, MRI. Recently, engineer and clinician apply real-time image navigation and image-guided surgery in the field of medical robotics surgery. In this report, we applied for obstetric diagnostic imaging to evaluate placenta accrete. Case: 42 years old, 29 weeks of gestation. 4G3P (3 times cesarean section, once abortion). She had suspected placenta accrete with placenta previa. We have been blood flow evaluation of the placenta attachment site on the uterine wall using 3D power-Doppler during hospitalization, and also MR image load to evaluation into 3D Slicer, In 33 weeks of gestation, She has taken cesarean
Abstracts / Placenta 35 (2014) A1eA23
O-026. PLACENTAL AMINO ACID TRANSPORTER FUNCTION ADAPTS TO MATERNAL AND FETAL NUTRIENT CONDITIONS Eiji Shibata David, J. Askew, Yukiyo Aiko, Chiharu Tomonaga, Satoshi Aramaki, Hirohide Inagaki, Toru Hachisuga. Department of Obstetrics and Gynecology, University of Occupational and Environmental Health, Japan Background: Studies suggest that placental nutrient uptake activity is responsive to both maternal and fetal nutrient demands. We hypothesize that under conditions of limited nutrient availability to the fetus, as often present in preeclampsia (PE), IUGR, and insufficient weight-gain during pregnancy, a general adaptive response aimed to increase amino acid transport activity may be observed in the placenta. Method: A total of 40 placentas from full-term (n¼10) and pre-term (n¼10) normal pregnancies, IUGR (n¼10), and PE (n¼10) associated pregnancies were looked at by immunohistochemistry followed by relative qualitative scoring to compare expression levels and localization of System L, ASCT2, and mTOR proteins. Result: Syncytiotrophoblast (ST) in placenta of pregnancies complicated by IUGR or PE showed significant increases in the levels of 4F2hc and LAT1 compared to both full-term control and pre-term pregnancies seperately. Elevated mTOR protein was uniquely higher in IUGR placentas compared to full-term controls. Total cellular ASCT2 transporter protein levels were similar in all groups, however, levels of ASCT2 protein localized to the ST microvillous membrane (MVM) were significantly lower in IUGR compared to both full-term and pre-term pregnancies. Additionally, ASCT2 and mTOR protein levels were positively associated with maternal prepregnancy BMI. Conclusion: First, in conditions of limited nutrient availability, such as PE or IUGR, there is an overall increase in the level of System L and mTOR protein expression in the ST, suggestive of an adaptive response. Second, a decrease in ASCT2 protein at the ST MVM suggests a post-translational event that may decrease AAT activity in IUGR placentas. Third, a physiological link between transporter expression and pre-pregnancy BMI is suggested based upon a positive association observed with ASCT2 and mTOR expression values.
O-027. ENDOMETRIAL PERIVASCULAR CELLS ESTABLISH UNIQUE CYTOKINE GRADIENTS IN DECIDUAL PROCESS a
a
a
Keisuke Murakami , Keiji Kuroda , Shintaro Makino , Atsuo Itakura a, Satoru Takeda a, Jan J. Brosens b. a Department of Obstetrics and Gynaecology, Juntendo University Faculty of Medicine, Japan; b Division of Reproductive Health, Clinical Science Research Laboratories, Warwick Medical School, University of Warwick, Japan Aim: The endometrial perivascular microenvironment is rich in mesenchymal stem-like cells that express type 1 integral membrane protein Sushi domain containing 2 (SUSD2) but the role of these cells in the decidual transformation of this tissue in pregnancy is unknown. We used an antibody directed against SUSD2 to isolate perivascular (SUSD2+) and non-perivascular (SUSD2-) fibroblasts from mid-luteal biopsies. Method: SUSD2+ and SUSD2- cells were purified from mid-luteal endometrial biopsies by magnetic bead separation and each cell population were expanded in vitro, then, subjected to RNA-seq analysis or differentiation experiments induced by cAMP and MPA to assess secretome profile. Result: On average, positive cells comprised 6.8% of endometrial stromal cells and 6-fold enriched in clonogenic cells. RNA sequencing revealed that cultures derived from SUSD2+ progenitor cells remain phenotypically distinct by the enrichment of novel and established endometrial perivascular signature genes including AOC3, NOTCH3, ELN, MYH11, DES, ACTA2, MCAM, and CSPG4. Decidualization transformed perivascular SUSD2+ -derived cells into the dominant source of a range of chemokines and cytokines, including CCL7, LIF and LTA.
Conclusion: Our findings confirmed that SUSD2+ -derived cells remain the expression of perivascular signature genes even after in-vitro culture. Decidual response is spatially organized at the feto-maternal interface with differentiating perivascular cells establishing distinct cytokine and chemokine gradients that could have the key role in decidual process.
O-028. LOCALIZATION OF TRPV5 VANILLOID RECEPTOR 5) TROPHOBLASTS
(TRANSIENT RECEPTOR PROTEIN IN HUMAN
POTENTIAL PLACENTAL
Yoko Aoyama, David J. Askew, Yukiyo Aiko, Chiharu Tomonaga, Satoshi Aramaki, Hirohide Inagaki, Toru Hachisuga, Eiji Shibata. Department of Obstetrics and Gynecology, University of Occupational and Environmental Health, Japan The Transient Receptor Proteins TRPV 5 and TRPV6 are members of a highly conserved family of Calcium channels sensitive to temperature, proteases, chemicals, shear stress, as well as intracellular Ca2+. TRPV 5 and 6 are both highly selective for Ca2+ as well as certain metal ions including Cadmium and Zinc. TRPV 5 and 6 are expressed in the placenta, and are thought to be the primary Ca2+ channels of the syncytiotroplast, and responsible for active Ca2+ transport across the placenta. Ca2+ transport is critical for proper fetal development, particularly bone calcification. Recent reports describe reduced activity and expression of these Ca2+ channels in the trophoblast of Preeclampsia-associated placenta. However, only TRPV6 has been shown to be expressed in the syncytiotrophoblast. Using immunohistochemistry, we found that TRPV 5 protein expression is primarily expressed in the cytotrophoblasts of the microvillous. TRPV5 staining in the syncytiotrophblast was very limited, primarily restricted to the nuclear syncytium. Therefore, the localization of TRPV5 with the placental microvillous is quite different from TRPV 6. In addition, our preliminary studies found no significant differences in TRPV 5 expression levels in the placenta of Preeclampsia or IUGR- associated pregnancies compared to controls. Our data suggests that TRPV5 and TRPV6 have distinct roles in the placenta.
O-029. THE NEW APPROACH TO DIAGNOSIS AND EVALUATE OF PLACENTA ACCRETE USING 3D SLICER Rie Oyama, Chizuko Isurugi, Shino Tanaka, Tomoyuki Fukagawa, Ikue Nakayama, Yuri Sasaki, Tomonobu Kanasugi, Aikihiko Kikuchi, Toru Sugiyama. Iwate Medical University Department Obstetrics and Gynecology, Japan Introduction: placenta accrete is the most important disease of the obstetric filed. In particular, in the case of a placenta that is attached over the side to posterior uterine in third trimester, it is necessary to evaluate the location of placenta sites, Therefore, we use various applications of ultrasound equipment, and combination with MR image. So, we tried to evaluate the identification of the attachment site of placenta accrete using 3D Slicer. 3D Slicer is open source -medical image software, which based at Harvard Medical School that consists of more than over 370k lines of code, mostly C++. This massive software development effort has been enabled by the participation of several large scale. NIH funded efforts, including the NA-MIC, NAC, BIRN, CIMIT and NCIGT communities (www.slicer.org). This software could able to evaluate the region of interest in the placenta attached uterine wall, and to create reconstructing image (for example: using affine transformation). 3D reconstructing image have processed on DICOM data (Raw volume data) of ultrasound, CT, MRI. Recently, engineer and clinician apply real-time image navigation and image-guided surgery in the field of medical robotics surgery. In this report, we applied for obstetric diagnostic imaging to evaluate placenta accrete. Case: 42 years old, 29 weeks of gestation. 4G3P (3 times cesarean section, once abortion). She had suspected placenta accrete with placenta previa. We have been blood flow evaluation of the placenta attachment site on the uterine wall using 3D power-Doppler during hospitalization, and also MR image load to evaluation into 3D Slicer, In 33 weeks of gestation, She has taken cesarean