P-350 Recombinant human anti-Müllerian hormone reduces doxorubicin-induced reproductive aging by regulating Id3 expression and promoting DNA repair
Abstract Study question What is the mechanism of ovarian damage after doxorubicin (Dox) treatment and how recombinant human anti-Müllerian hormone (rhAMH) suppresses ovarian aging and prevents chemotherapy-induced damage? Summary answer Administration of rhAMH attenuates Dox-induced follicle depleti...
Saved in:
Published in | Human reproduction (Oxford) Vol. 39; no. Supplement_1 |
---|---|
Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
03.07.2024
|
Online Access | Get full text |
Cover
Loading…
Abstract | Abstract Study question What is the mechanism of ovarian damage after doxorubicin (Dox) treatment and how recombinant human anti-Müllerian hormone (rhAMH) suppresses ovarian aging and prevents chemotherapy-induced damage? Summary answer Administration of rhAMH attenuates Dox-induced follicle depletion and protects the ovarian reserve in mice, primarily by reducing DNA damage via Id3 upregulation. What is known already Treatment with rhAMH results in reversible ovarian quiescence that can reduce primordial follicle depletion induced by Dox when it is used as an adjuvant. However, the exact mechanism of Dox-induced ovarian toxicity and the mechanisms by which rhAMH rescue this damage were unknown. Study design, size, duration For the in vivo experiments: 25-day-old mice were randomized into four treatment groups (Control, Dox, Dox+rhAMH, and rhAMH) for three different timepoints (n = 5 per group, total n = 60). Ovaries were removed for analysis at 4, 24 hours and and 1 week after treatment. For the in vitro studies: 21-day-old mice (n = 30) were treated with subcutaneous injection of diethylstilbestrol (DES) to enhance preantral follicle growth, and primary granulosa cells were collected by mechanical dissociation. Participants/materials, setting, methods Immunohistochemistry with DDX4 and counter staining with hematoxylin was used for differential follicle counts. DNA damage and apoptosis were measured by quantification of phosphorylated form of histone 2AX (γH2AX) and by TUNEL assay. MTS assay, clonogenic assay and siRNA Id3 RNA interference knockdown experiment were carried out to reveal the effects of doxorubicin and rhAMH on primary granulosa cell survival. Main results and the role of chance Treatment with Dox alone caused significant loss of PMF reserve (59% reduction, 5.25 versus 12.76 in control, P<0.0001) and this was significantly attenuated by coadministration with rhAMH (33% rescue, 9.47 P<0.001). The DNA damage marker γH2AX was elevated in primordial follicle (PMF) and primary follicle (PF) oocytes from Dox-treated ovaries 4 h after treatment, compared with controls (P<0.0001) and was significantly reduced (95% reduction) in rhAMH cotreated ovaries (P<0.001). Oocytes of secondary follicle (SF) and antral follicle (AF) were negative for γH2AX while granulosa cells (GCs) showed increased positivity for γH2AX after Dox treatment with significant reduction after rhAMH cotreatment. (24% reduction, P<0.001). Oocytes and somatic cells of PMF and PF rarely showed positive TUNEL staining at any of the timepoints evaluated.In vitro study showed reduced double stranded breaks (DSBs) in Dox-rhAMH cotreated GC compared to Dox alone indicated by number and signal intensity of γH2AX positive cells. Finally, while rhAMH treatment increased Id3 expression in early stage oocyte and granulosa cells, siRNA Id3 knock-down in granulosa cells significantly reduced the protective effect of rhAMH, as seen by the increased DSBs and pyknotic morphology of cells after Dox suggesting Id3 is downstream of AMH and enhancing Dox-induced DSB DNA repair. Limitations, reasons for caution This study failed to detect fate of PMF after DNA damage. PMF distinguished without apoptotic finding and this might be because experiment was conducted in limited time point or mechanism other than apoptotic cell death are responsible for PMF loss such as shedding, phagocytosis or necrosis. Wider implications of the findings Understanding of the mechanism behind rhAMH prevention of chemotherapy-induced reproductive aging is important for reproductive health and longevity. Possibility of rhAMH as non-invasive ferto-protective agent during chemotherapy may overcome limitation of current fertility preservation method which is limited by time requirement, cost and success rate. Trial registration number ’not applicable’ |
---|---|
AbstractList | Abstract Study question What is the mechanism of ovarian damage after doxorubicin (Dox) treatment and how recombinant human anti-Müllerian hormone (rhAMH) suppresses ovarian aging and prevents chemotherapy-induced damage? Summary answer Administration of rhAMH attenuates Dox-induced follicle depletion and protects the ovarian reserve in mice, primarily by reducing DNA damage via Id3 upregulation. What is known already Treatment with rhAMH results in reversible ovarian quiescence that can reduce primordial follicle depletion induced by Dox when it is used as an adjuvant. However, the exact mechanism of Dox-induced ovarian toxicity and the mechanisms by which rhAMH rescue this damage were unknown. Study design, size, duration For the in vivo experiments: 25-day-old mice were randomized into four treatment groups (Control, Dox, Dox+rhAMH, and rhAMH) for three different timepoints (n = 5 per group, total n = 60). Ovaries were removed for analysis at 4, 24 hours and and 1 week after treatment. For the in vitro studies: 21-day-old mice (n = 30) were treated with subcutaneous injection of diethylstilbestrol (DES) to enhance preantral follicle growth, and primary granulosa cells were collected by mechanical dissociation. Participants/materials, setting, methods Immunohistochemistry with DDX4 and counter staining with hematoxylin was used for differential follicle counts. DNA damage and apoptosis were measured by quantification of phosphorylated form of histone 2AX (γH2AX) and by TUNEL assay. MTS assay, clonogenic assay and siRNA Id3 RNA interference knockdown experiment were carried out to reveal the effects of doxorubicin and rhAMH on primary granulosa cell survival. Main results and the role of chance Treatment with Dox alone caused significant loss of PMF reserve (59% reduction, 5.25 versus 12.76 in control, P<0.0001) and this was significantly attenuated by coadministration with rhAMH (33% rescue, 9.47 P<0.001). The DNA damage marker γH2AX was elevated in primordial follicle (PMF) and primary follicle (PF) oocytes from Dox-treated ovaries 4 h after treatment, compared with controls (P<0.0001) and was significantly reduced (95% reduction) in rhAMH cotreated ovaries (P<0.001). Oocytes of secondary follicle (SF) and antral follicle (AF) were negative for γH2AX while granulosa cells (GCs) showed increased positivity for γH2AX after Dox treatment with significant reduction after rhAMH cotreatment. (24% reduction, P<0.001). Oocytes and somatic cells of PMF and PF rarely showed positive TUNEL staining at any of the timepoints evaluated.In vitro study showed reduced double stranded breaks (DSBs) in Dox-rhAMH cotreated GC compared to Dox alone indicated by number and signal intensity of γH2AX positive cells. Finally, while rhAMH treatment increased Id3 expression in early stage oocyte and granulosa cells, siRNA Id3 knock-down in granulosa cells significantly reduced the protective effect of rhAMH, as seen by the increased DSBs and pyknotic morphology of cells after Dox suggesting Id3 is downstream of AMH and enhancing Dox-induced DSB DNA repair. Limitations, reasons for caution This study failed to detect fate of PMF after DNA damage. PMF distinguished without apoptotic finding and this might be because experiment was conducted in limited time point or mechanism other than apoptotic cell death are responsible for PMF loss such as shedding, phagocytosis or necrosis. Wider implications of the findings Understanding of the mechanism behind rhAMH prevention of chemotherapy-induced reproductive aging is important for reproductive health and longevity. Possibility of rhAMH as non-invasive ferto-protective agent during chemotherapy may overcome limitation of current fertility preservation method which is limited by time requirement, cost and success rate. Trial registration number ’not applicable’ |
Author | Chauvin, M Kashiwagi, A May, P Chang, E M Pepin, D Godin, P Nagykery, N |
Author_xml | – sequence: 1 givenname: E M surname: Chang fullname: Chang, E M – sequence: 2 givenname: P surname: Godin fullname: Godin, P – sequence: 3 givenname: P surname: May fullname: May, P – sequence: 4 givenname: M surname: Chauvin fullname: Chauvin, M – sequence: 5 givenname: A surname: Kashiwagi fullname: Kashiwagi, A – sequence: 6 givenname: N surname: Nagykery fullname: Nagykery, N – sequence: 7 givenname: D surname: Pepin fullname: Pepin, D |
BookMark | eNqdj01OwzAQhS1UJFrgAqx8Abd2AiEsET-CBQgh9pYTT9NBiR2NE9Teg-Ow42JMoCdgNW_emx99CzELMYAQZ0Yvjb7KV5uxI-hXHhwYXS71pTkQc3NeaJXlF3om5jorSmVMYY7EIqV3rVmWxVx8vigekK9Qx67C4MIg-ZYLkhWqp--vtgVC7jeROv4pCfxYQ5I-biONFdYYFIbJ85z1FFkO-AHSNRgaWe3YbcbWDVP36HMJ254gJYzTDy95o4u_4e3z9XTBIZ2Iw7VrE5zu67HI7u_ebh5UTTElgrXtCTtHO2u0nfjtH7_d81vmz_-19AMgYG4r |
ContentType | Journal Article |
DBID | AAYXX CITATION |
DOI | 10.1093/humrep/deae108.071 |
DatabaseName | CrossRef |
DatabaseTitle | CrossRef |
DatabaseTitleList | CrossRef |
DeliveryMethod | fulltext_linktorsrc |
Discipline | Medicine Anatomy & Physiology Pharmacy, Therapeutics, & Pharmacology |
EISSN | 1460-2350 |
ExternalDocumentID | 10_1093_humrep_deae108_071 |
GroupedDBID | --- -E4 .2P .I3 .XZ .ZR 0R~ 1TH 29I 2WC 4.4 482 48X 53G 5GY 5RE 5VS 5WA 5WD 70D AABZA AACZT AAIMJ AAJKP AAMDB AAMVS AAOGV AAPNW AAPQZ AAPXW AARHZ AASNB AAUAY AAVAP AAVLN AAYXX ABEUO ABIXL ABJNI ABKDP ABMNT ABNHQ ABNKS ABPTD ABQLI ABQNK ABWST ABXVV ABZBJ ACCCW ACGFS ACPRK ACUFI ACUTO ACYHN ADBBV ADEYI ADEZT ADGKP ADGZP ADHKW ADHZD ADIPN ADJQC ADOCK ADQBN ADRIX ADRTK ADVEK ADYVW ADZXQ AEGPL AEJOX AEKSI AELWJ AEMDU AENEX AENZO AEPUE AETBJ AEWNT AFFZL AFGWE AFIYH AFOFC AFXAL AFXEN AGINJ AGKEF AGQXC AGSYK AGUTN AHMBA AHXPO AIJHB AJEEA AKWXX ALMA_UNASSIGNED_HOLDINGS ALUQC APIBT APWMN ARIXL ATGXG AXUDD AYOIW BAWUL BAYMD BCRHZ BEYMZ BHONS BQDIO BSWAC BTRTY BVRKM C45 CDBKE CITATION CS3 CZ4 DAKXR DILTD DU5 D~K E3Z EBS EE~ ENERS F5P F9B FECEO FHSFR FLUFQ FOEOM FOTVD FQBLK GAUVT GJXCC H13 H5~ HAR HW0 HZ~ IOX J21 JXSIZ KAQDR KOP KQ8 KSI KSN L7B M-Z MHKGH N9A NGC NLBLG NOMLY NOYVH O9- OAUYM OAWHX OBOKY OCZFY ODMLO OJQWA OJZSN OK1 OPAEJ OVD OWPYF P2P PAFKI PEELM PQQKQ Q1. Q5Y R44 RD5 RIG ROL ROX ROZ RUSNO RW1 RXO TCURE TEORI TJX TLC TR2 WH7 WOQ X7H YAYTL YKOAZ YXANX ZKX ~91 |
ID | FETCH-crossref_primary_10_1093_humrep_deae108_0713 |
ISSN | 0268-1161 |
IngestDate | Thu Sep 12 20:56:04 EDT 2024 |
IsPeerReviewed | true |
IsScholarly | true |
Issue | Supplement_1 |
Language | English |
LinkModel | OpenURL |
MergedId | FETCHMERGED-crossref_primary_10_1093_humrep_deae108_0713 |
ParticipantIDs | crossref_primary_10_1093_humrep_deae108_071 |
PublicationCentury | 2000 |
PublicationDate | 2024-07-03 |
PublicationDateYYYYMMDD | 2024-07-03 |
PublicationDate_xml | – month: 07 year: 2024 text: 2024-07-03 day: 03 |
PublicationDecade | 2020 |
PublicationTitle | Human reproduction (Oxford) |
PublicationYear | 2024 |
SSID | ssj0016186 |
Score | 4.978372 |
Snippet | Abstract Study question What is the mechanism of ovarian damage after doxorubicin (Dox) treatment and how recombinant human anti-Müllerian hormone (rhAMH)... |
SourceID | crossref |
SourceType | Aggregation Database |
Title | P-350 Recombinant human anti-Müllerian hormone reduces doxorubicin-induced reproductive aging by regulating Id3 expression and promoting DNA repair |
Volume | 39 |
hasFullText | 1 |
inHoldings | 1 |
isFullTextHit | |
isPrint | |
link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV1Nb9NAEF1FRaq4VJCAaKHVHJpeWrf-yId7TENQBHIUpCD1ZtlZR-RQG5m4qvkd_BVuvfWPdWZnYy9VkEgvluPYE9vzsju78_aNEMeX2GvFDim3dnu-RZVgLR_bYsuNXGfev-zKSKVigklv_K3z-bp73Wj8MVhLxSo-n__auK7kOV7FY-hXWiW7hWcro3gA99G_uEUP4_a_fDy1vK5NkV92EytGiy65h3tLK6Ac-NWQlvrRn_g7BqdZSkVSZEEsLJndZXkRU2LdwnF5QTwAUrhUArBEJ-LyRRid5lyuXlELpEc1AZg7m2qRAcXnwy8_TgZkIVrmZsTLWYLaskKbz-sUjWmI4XrienTaHrrtgW0o90vWOahWogVR-ddnvLa45XMCcxbD7SjGq2c0di5CxXFYmP084ca407Mt12Nh2nVrzdJHGpWq8qmaRg2djf0Ba2Xhu8fHxB2ZRIlD7Nm-U_d_65z_k26xIitymt4L2UqobYQ2KRe8cBHPNOb_8rVOXlEJAp7a40fSa7XQxgXbuDDuw4iHjMBm9krs6REJDBher0UjSZuiNUijVXZTwgkojrBKvjTFbqCpGE1xMmXR8_IMZvUavp9n6opKDr1sid8KpWCgFBRKgVH6cK8RChqhoBEKGxAKJkJBIRTiEmqEAiIUaoTib0ioEAqIUGCEvhHup9FsOLbWbyX8wbIr4b_94L0VOyne4DsB0vd97L8j6S9w7G8v4r6ziOxkbi866Kmkty9OtzB8sNXZ78XLGtofxM4qL5JDDFlX8ZECyCMwY6NA |
link.rule.ids | 315,786,790,27957,27958 |
linkProvider | Colorado Alliance of Research Libraries |
openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=P-350+Recombinant+human+anti-M%C3%BCllerian+hormone+reduces+doxorubicin-induced+reproductive+aging+by+regulating+Id3+expression+and+promoting+DNA+repair&rft.jtitle=Human+reproduction+%28Oxford%29&rft.au=Chang%2C+E+%C2%A0M&rft.au=Godin%2C+P&rft.au=May%2C+P&rft.au=Chauvin%2C+M&rft.date=2024-07-03&rft.issn=0268-1161&rft.eissn=1460-2350&rft.volume=39&rft.issue=Supplement_1&rft_id=info:doi/10.1093%2Fhumrep%2Fdeae108.071&rft.externalDBID=n%2Fa&rft.externalDocID=10_1093_humrep_deae108_071 |
thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=0268-1161&client=summon |
thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=0268-1161&client=summon |
thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=0268-1161&client=summon |