RODRÍGUEZ LOMBARDI GONZALO RAMÓN
Congresos y reuniones científicas
Título:
REGULATORY TRANSCRIPTIONAL DOMAIN OF KRÜPPEL-LIKE FACTOR 6 (KLF6) IS NECESSARY FOR PLACENTAL TROPHOBLAST CELLS FUSION
Lugar:
Mar del Plata
Reunión:
Congreso; Reunión Conjunta de Sociedades SAIC-SAI-SAFIS; 2019
Institución organizadora:
SAIC-SAI-SAFIS
Resumen:
Placental development and maintenance require the fusion of villous cytotrophoblasts
(vCTB) to generate the syncytiotrophoblast layer involved in the placental barrier function,
synthesis of critical proteins and nutrient exchange between the mother and the fetus. The
Krüppel-like factor 6 (KLF6) is highly expressed in placenta and Klf6-/- mice die at day
E12.5 showing impaired placenta development. Previously, we have demonstrated that
KLF6 modulates the expression of the cell cycle inhibitory protein p21, of Syn-1, and β-
hCG, and it is required for cell?cell fusion in human primary vCTB as well as in the BeWo
trophoblast-derived cell line. More importantly, KLF6 is sufficient to trigger cell fusion in
BeWo cells. Herein we analysed the KLF6-dependent mechanisms involved in KLF6-
induced fusion. Stable BeWo cell lines containing a KLF6 mutant that lacks the acidic
domain required for its transcriptional activity (KLF6∆ac-BeWo), the full-length KLF6
protein (KLF6-BeWo), or the Empty-vector (EV-BeWo) were generated. KLF6∆ac-BeWo
cells were treated or not with 30 μM forskolin and BeWo-KLF6 cells were transduced with
KLF6∆ac lentivirus particles and cell fusion was analysed by immunofluorescence after 72
h. Transcript and/or protein levels of Syn-1, β-hCG, Cx43, ABCG2, Gal-1, and GRP78
were measured by qRT-PCR and western blot, respectively. KLF6-BeWo cells were treated
with a specific p21 siRNA or a scramble siRNA as control. KLF6 overexpression induced
the formation of syncytial structures and increased the expression of molecules involved in
this process. While, KLF6∆ac did not promote syncytialization, impaired forskolin-induced
cell fusion, and decreased Syn-1 and β-hCG expression. Moreover, cell fusion was reduced
in KLF6-BeWo cells transduced with the deletion mutant lentivirus or silenced for p21.
These results suggest that KLF6 induces vCTB syncytialization through a mechanism that
involves its regulatory transcriptional domain in a p21-dependent manner.
(vCTB) to generate the syncytiotrophoblast layer involved in the placental barrier function,
synthesis of critical proteins and nutrient exchange between the mother and the fetus. The
Krüppel-like factor 6 (KLF6) is highly expressed in placenta and Klf6-/- mice die at day
E12.5 showing impaired placenta development. Previously, we have demonstrated that
KLF6 modulates the expression of the cell cycle inhibitory protein p21, of Syn-1, and β-
hCG, and it is required for cell?cell fusion in human primary vCTB as well as in the BeWo
trophoblast-derived cell line. More importantly, KLF6 is sufficient to trigger cell fusion in
BeWo cells. Herein we analysed the KLF6-dependent mechanisms involved in KLF6-
induced fusion. Stable BeWo cell lines containing a KLF6 mutant that lacks the acidic
domain required for its transcriptional activity (KLF6∆ac-BeWo), the full-length KLF6
protein (KLF6-BeWo), or the Empty-vector (EV-BeWo) were generated. KLF6∆ac-BeWo
cells were treated or not with 30 μM forskolin and BeWo-KLF6 cells were transduced with
KLF6∆ac lentivirus particles and cell fusion was analysed by immunofluorescence after 72
h. Transcript and/or protein levels of Syn-1, β-hCG, Cx43, ABCG2, Gal-1, and GRP78
were measured by qRT-PCR and western blot, respectively. KLF6-BeWo cells were treated
with a specific p21 siRNA or a scramble siRNA as control. KLF6 overexpression induced
the formation of syncytial structures and increased the expression of molecules involved in
this process. While, KLF6∆ac did not promote syncytialization, impaired forskolin-induced
cell fusion, and decreased Syn-1 and β-hCG expression. Moreover, cell fusion was reduced
in KLF6-BeWo cells transduced with the deletion mutant lentivirus or silenced for p21.
These results suggest that KLF6 induces vCTB syncytialization through a mechanism that
involves its regulatory transcriptional domain in a p21-dependent manner.
