1/20
New in @ATSBlueEditor, the culmination of over 4 years of work in the @bradleymaron lab, in which we identify that #NEDD9 is a hypoxia-regulated pulmonary endothelial protein that directly binds activated platelets to promote pulmonary thrombosis.
atsjournals.org/doi/10.1164/rc…
New in @ATSBlueEditor, the culmination of over 4 years of work in the @bradleymaron lab, in which we identify that #NEDD9 is a hypoxia-regulated pulmonary endothelial protein that directly binds activated platelets to promote pulmonary thrombosis.
atsjournals.org/doi/10.1164/rc…
2/20
The @bradleymaron lab previously discovered that the malignancy protein #NEDD9 promotes endothelial fibrosis by regulating collagen synthesis in human pulmonary artery endothelial cells (HPAECs), which could be targeted to inhibit #PAH in vivo.
stm.sciencemag.org/content/10/445…
The @bradleymaron lab previously discovered that the malignancy protein #NEDD9 promotes endothelial fibrosis by regulating collagen synthesis in human pulmonary artery endothelial cells (HPAECs), which could be targeted to inhibit #PAH in vivo.
stm.sciencemag.org/content/10/445…
3/20
As a pro-metastatic protein, #NEDD9 interacts with pro-thrombotic cell types, & it was shown to be ⬆️ via #HIF1α in cancer cells to promote migration. We hypothesized that #NEDD9 is ⬆️ by hypoxia in HPAECs ➡️ pathogenic platelet-EC adhesion.
cancerres.aacrjournals.org/content/70/10/…
As a pro-metastatic protein, #NEDD9 interacts with pro-thrombotic cell types, & it was shown to be ⬆️ via #HIF1α in cancer cells to promote migration. We hypothesized that #NEDD9 is ⬆️ by hypoxia in HPAECs ➡️ pathogenic platelet-EC adhesion.
cancerres.aacrjournals.org/content/70/10/…
5/20
We performed a network analysis using hypoxia-regulated transcripts from HPAEC RNA-Seq enriched for thrombosis genes mapped to the human interactome. #NEDD9 emerged in the "hypoxia-thrombosome," providing evidence in silico that HPAEC #NEDD9 may be relevant to thrombosis.
We performed a network analysis using hypoxia-regulated transcripts from HPAEC RNA-Seq enriched for thrombosis genes mapped to the human interactome. #NEDD9 emerged in the "hypoxia-thrombosome," providing evidence in silico that HPAEC #NEDD9 may be relevant to thrombosis.
6/20
Immunofluorescence of non-permeabilized HPAECs showed #NEDD9 was ⬆️ by hypoxia & co-localized with the endothelial plasma membrane protein #PECAM1. Plasma membrane localization was further confirmed by immunoblot & LC-MS of HPAEC plasma membrane isolates.
Immunofluorescence of non-permeabilized HPAECs showed #NEDD9 was ⬆️ by hypoxia & co-localized with the endothelial plasma membrane protein #PECAM1. Plasma membrane localization was further confirmed by immunoblot & LC-MS of HPAEC plasma membrane isolates.
7/20
When we inhibited HPAEC #NEDD9 using siRNA, we demonstrated a significant decrease in platelet-HPAEC adhesion under conditions of platelet activation with thrombin receptor agonist peptide (#TRAP), suggesting #NEDD9 interacts with activated platelets.
When we inhibited HPAEC #NEDD9 using siRNA, we demonstrated a significant decrease in platelet-HPAEC adhesion under conditions of platelet activation with thrombin receptor agonist peptide (#TRAP), suggesting #NEDD9 interacts with activated platelets.
8/20
Using immunoprecipitation, LC-MS, & microscale thermophoresis, we identified the substrate domain of #NEDD9 binds directly to #PSelectin. We then generated a custom-made monospecific polyclonal antibody (msAb-N9) targeting the putative binding region b/w #NEDD9-#PSelectin.
Using immunoprecipitation, LC-MS, & microscale thermophoresis, we identified the substrate domain of #NEDD9 binds directly to #PSelectin. We then generated a custom-made monospecific polyclonal antibody (msAb-N9) targeting the putative binding region b/w #NEDD9-#PSelectin.
9/20
Using custom msAb-N9, we showed that we could significantly decrease adhesion between HPAECs and #TRAP-activated platelets, suggesting the #NEDD9-#PSelectin binding interaction was important for platelet-endothelial interactions, at least in vitro.
Using custom msAb-N9, we showed that we could significantly decrease adhesion between HPAECs and #TRAP-activated platelets, suggesting the #NEDD9-#PSelectin binding interaction was important for platelet-endothelial interactions, at least in vitro.
10/20
In a model of acute platelet activation with #ADP, msAb-N9-treated mice had ⬇️ pulmonary thrombotic occlusion & #PH in vivo, akin to the #NEDD9KO mice. In a model of chronic thromboembolic remodeling, msAb-N9-treated rats had ⬇️ platelet aggregates & vascular remodeling.
In a model of acute platelet activation with #ADP, msAb-N9-treated mice had ⬇️ pulmonary thrombotic occlusion & #PH in vivo, akin to the #NEDD9KO mice. In a model of chronic thromboembolic remodeling, msAb-N9-treated rats had ⬇️ platelet aggregates & vascular remodeling.
11/20
Compared to control & acute #DVT / #PE, patients with #CTEPH had ⬆️ #NEDD9 & #HIF1α expression. Thanks to @SudarRajagopal et al., we show #CTEPH PAECs also had ⬆️ #NEDD9 & #HIF1α, & msAb-N9 ⬇️ platelet-#CTEPH PAEC adhesion ex vivo. #CTEPH patients had ⬆️ plasma #NEDD9.
Compared to control & acute #DVT / #PE, patients with #CTEPH had ⬆️ #NEDD9 & #HIF1α expression. Thanks to @SudarRajagopal et al., we show #CTEPH PAECs also had ⬆️ #NEDD9 & #HIF1α, & msAb-N9 ⬇️ platelet-#CTEPH PAEC adhesion ex vivo. #CTEPH patients had ⬆️ plasma #NEDD9.
12/20
In sum, we identified a novel molecular target of activated platelets with mechanistic and pharmacotherapeutic implications for #CTEPH (among other pulmonary vascular thrombotic disorders).
In sum, we identified a novel molecular target of activated platelets with mechanistic and pharmacotherapeutic implications for #CTEPH (among other pulmonary vascular thrombotic disorders).
13/20
I will conclude by saying: choosing a postdoctoral research pathway was NOT EASY for me. I had an interest in #CTEPH pathobiology & a desire to "identify novel therapeutic targets," but I had not set foot in a lab in almost a decade. Many laughed me off. Not @BradleyMaron.
I will conclude by saying: choosing a postdoctoral research pathway was NOT EASY for me. I had an interest in #CTEPH pathobiology & a desire to "identify novel therapeutic targets," but I had not set foot in a lab in almost a decade. Many laughed me off. Not @BradleyMaron.
14/20
Within a few months of working with @BradleyMaron I had enough of my own preliminary data to secure a #NRSA #F32 via @nih_nhlbi. It was his #mentorship and #sponsorship that laid the groundwork for the success of this project and my subsequent career development.
Within a few months of working with @BradleyMaron I had enough of my own preliminary data to secure a #NRSA #F32 via @nih_nhlbi. It was his #mentorship and #sponsorship that laid the groundwork for the success of this project and my subsequent career development.
15/20
Thank you to all the support I have received from @nih_nhlbi, @NIH_LRP, @HarvardCatalyst, @BBIC_NCAI, @AHAScience, @atscommunity, @HarvardPulm, @MGHMedicine, & @MGH_RI, along w/ tremendous collaborators @BradWertheim @SudarRajagopal @rchannick #JoeLoscalzo #BethBattinelli
Thank you to all the support I have received from @nih_nhlbi, @NIH_LRP, @HarvardCatalyst, @BBIC_NCAI, @AHAScience, @atscommunity, @HarvardPulm, @MGHMedicine, & @MGH_RI, along w/ tremendous collaborators @BradWertheim @SudarRajagopal @rchannick #JoeLoscalzo #BethBattinelli
16/20
Special shout out to @BurnettBowie who was the first person to tell me "yes, you CAN do basic science," & whose sage counsel has guided me since I walked in the door @MGHMedicine. Huge thanks to @mghcdi @MGHEquity for the support & working to ⬆️ URiM physician-scientists.
Special shout out to @BurnettBowie who was the first person to tell me "yes, you CAN do basic science," & whose sage counsel has guided me since I walked in the door @MGHMedicine. Huge thanks to @mghcdi @MGHEquity for the support & working to ⬆️ URiM physician-scientists.
17/20
Thanks to peer mentors like @jehanalladina @BizarMd @JSawallaGusehMD @NnekaUfereMD @mbake013 @rsknipe @ChanaSacks @noshreza @DrAMcCullough for helping me navigate the ⬆️⬇️⏬🔂 of academia and whose careers I love to watch ⏫.
Thanks to peer mentors like @jehanalladina @BizarMd @JSawallaGusehMD @NnekaUfereMD @mbake013 @rsknipe @ChanaSacks @noshreza @DrAMcCullough for helping me navigate the ⬆️⬇️⏬🔂 of academia and whose careers I love to watch ⏫.
18/20
Thanks to the best partner in life anyone can ask for, @LucySchulson, who has had to deal with me coming home on weekend nights after moments like this 👇
Thanks to the best partner in life anyone can ask for, @LucySchulson, who has had to deal with me coming home on weekend nights after moments like this 👇
https://twitter.com/galbamd/status/1122559831880208386?s=20
19/20
One penultimate comment: this year was hard.
Notice the gap between receipt & acceptance. Anything happen during that time?
For all who struggled, contemplated quitting academia, & went through untold personal struggles while navigating [gestures broadly], I see you.
One penultimate comment: this year was hard.
Notice the gap between receipt & acceptance. Anything happen during that time?
For all who struggled, contemplated quitting academia, & went through untold personal struggles while navigating [gestures broadly], I see you.
20/20
I will leave you with some wise words from the best mentor that I could have asked for:
I will leave you with some wise words from the best mentor that I could have asked for:
https://twitter.com/BradleyMaron/status/1342937763067424769?s=20
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