This preprint is under consideration at Cancer Cell International. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
A new preprint design is coming!
We have improved readability, clarity, accessibility, and opportunities for engagement.
Primary research
Youxin Zhou
First Affiliated Hospital of Soochow University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-2835-8376
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Glioblastomas (GBMs) are grade IV central nervous system tumors characterized by a poor prognosis and a median overall survival of 15 months. The glioblastoma cell population is more genetically unstable, resistant to chemotherapy, more angiogenic, and more malignant. Because ubiquitin-specific proteases (USPs) remove ubiquitin from oncogenic protein substrates, effective inhibition of ubiquitin-specific protease 7 (USP7), which is highly expressed in GBMs, is a potentially critical therapeutic approach.
Methods: Immunohistochemistry and Western blotting were used to detect high expression of USP7 in GBM. in vitro studies were performed by Western blotting, immunofluorescence, and flow cytometry to detect apoptosis following inhibition of USP7. Anti-apoptotic substrates of USP7 were defined by Co-IP and TMT proteomics. Nude mouse intracranial xenograft models were constructed to verify whether inhibition of USP7 inhibited the proliferation rate of tumours.
Results: USP7 is significantly upregulated in glioblastoma samples. Interfering with USP7 in GBMs induced significant apoptosis, which also occurred after treatment with P5091, a novel small molecule inhibitor of USP7. Mechanistically, apoptosis of GBMs after interference with USP7 function is achieved by stabilizing a key anti-apoptotic protein, ADP-ribosylation factor 4 (ARF4).Furthermore, USP7 interacts directly with ARF4 and catalyzes the removal of the K48-linked polyubiquitinated chain that binds to ARF4, thereby stabilizing the protein. In in vivo experiments, P5091 significantly inhibited tumor growth and promoted the expression of apoptotic genes.
Conclusion: Targeted inhibition of USP7 enhances the ubiquitination of ARF4 and ultimately mediates the apoptosis of GBM cells. In a clinical sense, P5091 as a specific inhibitor of USP7 may be an effective approach for the treatment of GBM.
Keywords
GBM, USP7, Ubiquitination, ARF4, Apoptosis
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Loading...
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 01 Jun, 2021
No community comments so far
Editorial decision: Major revision
On 04 Jul, 2021
Review #11 received
Received 27 Jun, 2021
Review #4 received
Received 27 Jun, 2021
Review #8 received
Received 15 Jun, 2021
Review #5 received
Received 15 Jun, 2021
Review #7 received
Received 14 Jun, 2021
Review #6 received
Received 14 Jun, 2021
Reviewer #7 agreed
On 13 Jun, 2021
Reviewer #8 agreed
On 13 Jun, 2021
Reviewer #9 agreed
On 13 Jun, 2021
Reviewer #10 agreed
On 13 Jun, 2021
Reviewer #11 agreed
On 13 Jun, 2021
Reviewer #12 agreed
On 13 Jun, 2021
Reviewer #6 agreed
On 13 Jun, 2021
Review #3 received
Received 13 Jun, 2021
Reviewer #5 agreed
On 12 Jun, 2021
Reviewer #4 agreed
On 06 Jun, 2021
Reviewer #3 agreed
On 01 Jun, 2021
Reviews received
Received 01 Jun, 2021
Reviewer #2 agreed
On 31 May, 2021
Reviewers invited
Invitations sent on 31 May, 2021
Reviewer #1 agreed
On 31 May, 2021
Review #2 received
Received 31 May, 2021
Review #1 received
Received 31 May, 2021
Editor invited
On 25 May, 2021
Editor assigned
On 23 May, 2021
Submission checks complete
On 23 May, 2021
First submitted
On 22 May, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Cancer Biology
Learn more about our company.
A new preprint design is coming!
We have improved readability, clarity, accessibility, and opportunities for engagement.
This preprint is under consideration at Cancer Cell International. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Primary research
Youxin Zhou
First Affiliated Hospital of Soochow University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-2835-8376
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 01 Jun, 2021
No community comments so far
Editorial decision: Major revision
On 04 Jul, 2021
Review #11 received
Received 27 Jun, 2021
Review #4 received
Received 27 Jun, 2021
Review #8 received
Received 15 Jun, 2021
Review #5 received
Received 15 Jun, 2021
Review #7 received
Received 14 Jun, 2021
Review #6 received
Received 14 Jun, 2021
Reviewer #7 agreed
On 13 Jun, 2021
Reviewer #8 agreed
On 13 Jun, 2021
Reviewer #9 agreed
On 13 Jun, 2021
Reviewer #10 agreed
On 13 Jun, 2021
Reviewer #11 agreed
On 13 Jun, 2021
Reviewer #12 agreed
On 13 Jun, 2021
Reviewer #6 agreed
On 13 Jun, 2021
Review #3 received
Received 13 Jun, 2021
Reviewer #5 agreed
On 12 Jun, 2021
Reviewer #4 agreed
On 06 Jun, 2021
Reviewer #3 agreed
On 01 Jun, 2021
Reviews received
Received 01 Jun, 2021
Reviewer #2 agreed
On 31 May, 2021
Reviewers invited
Invitations sent on 31 May, 2021
Reviewer #1 agreed
On 31 May, 2021
Review #2 received
Received 31 May, 2021
Review #1 received
Received 31 May, 2021
Editor invited
On 25 May, 2021
Editor assigned
On 23 May, 2021
Submission checks complete
On 23 May, 2021
First submitted
On 22 May, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Cancer Biology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Glioblastomas (GBMs) are grade IV central nervous system tumors characterized by a poor prognosis and a median overall survival of 15 months. The glioblastoma cell population is more genetically unstable, resistant to chemotherapy, more angiogenic, and more malignant. Because ubiquitin-specific proteases (USPs) remove ubiquitin from oncogenic protein substrates, effective inhibition of ubiquitin-specific protease 7 (USP7), which is highly expressed in GBMs, is a potentially critical therapeutic approach.
Methods: Immunohistochemistry and Western blotting were used to detect high expression of USP7 in GBM. in vitro studies were performed by Western blotting, immunofluorescence, and flow cytometry to detect apoptosis following inhibition of USP7. Anti-apoptotic substrates of USP7 were defined by Co-IP and TMT proteomics. Nude mouse intracranial xenograft models were constructed to verify whether inhibition of USP7 inhibited the proliferation rate of tumours.
Results: USP7 is significantly upregulated in glioblastoma samples. Interfering with USP7 in GBMs induced significant apoptosis, which also occurred after treatment with P5091, a novel small molecule inhibitor of USP7. Mechanistically, apoptosis of GBMs after interference with USP7 function is achieved by stabilizing a key anti-apoptotic protein, ADP-ribosylation factor 4 (ARF4).Furthermore, USP7 interacts directly with ARF4 and catalyzes the removal of the K48-linked polyubiquitinated chain that binds to ARF4, thereby stabilizing the protein. In in vivo experiments, P5091 significantly inhibited tumor growth and promoted the expression of apoptotic genes.
Conclusion: Targeted inhibition of USP7 enhances the ubiquitination of ARF4 and ultimately mediates the apoptosis of GBM cells. In a clinical sense, P5091 as a specific inhibitor of USP7 may be an effective approach for the treatment of GBM.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Loading...