Bob Garcea

Phosphorylation of Human Papillomavirus Type 16 L2 Contributes to Efficient Virus Infectious Entry

Anonymous — Wed, 23 Oct 2019 17:22:22 +0000

Phosphorylation of Human Papillomavirus Type 16 L2 Contributes to Efficient Virus Infectious Entry Anonymous (not verified) Wed, 10/23/2019 - 11:22

The human papillomavirus (HPV) capsid comprises two viral proteins, L1 and L2, with the L2 component being essential to ensure efficient endocytic transport of incoming viral genomes. Several studies have previously reported that L1 and L2 are posttranslationally modified, but it is uncertain whether these modifications affect HPV infectious entry. Using a proteomic screen, we identified a highly conserved phospho-acceptor site on the HPV-16 and bovine papillomavirus 1 (BPV-1) L2 proteins. The phospho-modification of L2 and its presence in HPV pseudovirions (PsVs) were confirmed using anti-phospho-L2-specific antibodies. Mutation of the phospho-acceptor sites of both HPV-16 and BPV-1 L2 resulted in the production of infectious virus particles, with no differences in efficiencies of packaging the reporter DNA. However, these mutated PsVs showed marked defects in infectious entry. Further analysis revealed a defect in uncoating, characterized by a delay in the exposure of a conformational epitope on L1 that indicates capsid uncoating. This uncoating defect was accompanied by a delay in the proteolysis of both L1 and L2 in mutated HPV-16 PsVs. Taken together, these studies indicate that phosphorylation of L2 during virus assembly plays an important role in optimal uncoating of virions during infection, suggesting that phosphorylation of the viral capsid proteins contributes to infectious entry.

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Viral replication centers and the DNA damage response in JC virus-infected cells.

Anonymous — Wed, 23 Oct 2019 17:20:07 +0000

Viral replication centers and the DNA damage response in JC virus-infected cells. Anonymous (not verified) Wed, 10/23/2019 - 11:20

JCV is a human polyomavirus (PyV) that establishes a persistent infection in its host. Current immunomodulatory therapies, such as Natalizumab for multiple sclerosis, can result in JCV reactivation, leading to the debilitating brain disease progressive multifocal leukoencephalopathy (PML). JCV is among the viruses that recruit and modulate the host DNA damage response (DDR) to replicate its genome. We have identified host proteins recruited to the nuclear sites of JC viral DNA (vDNA) replication using three cell types susceptible to infection in vitro. Using confocal microscopy, we found that JCV recruited a similar repertoire of host DDR proteins to these replication sites previously observed for other PyVs. Electron tomography of JCV "virus factories" showed structural features like those described for murine PyV. These results confirm and extend previous observations for PyVs to JCV emphasizing a similar replication strategy among members of this virus family.

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$1.1 million grant funds CU �鶹ӰԺ research into next-generation vaccines

Anonymous — Fri, 04 Nov 2016 06:00:00 +0000

$1.1 million grant funds CU �鶹ӰԺ research into next-generation vaccines Anonymous (not verified) Fri, 11/04/2016 - 00:00

BioFrontiers

The �鶹ӰԺ has received a $1.1 million grant from the Bill & Melinda Gates Foundation to develop next-generation vaccines that require no refrigeration and defend against infectious diseases with just one shot.

If successful, those advancements could radically transform the difficult task of dispensing life-saving immunizations in developing countries — and improve convenience in every part of the world.

Professor Bob Garcea of the Department of Molecular, Cellular and Developmental Biology and the BioFrontiers Institute has teamed up with Professors Ted Randolph and Al Weimer of the Department of Chemical and Biological Engineering in a unique collaboration that applies a wide range of skillsets and ideas to the pressing challenge of delivering vaccines to patients in developing countries. All three investigators work in the Jennie Smoly Caruthers Biotechnology Building (JSCBB) at CU �鶹ӰԺ, but their research areas have very different emphases.

“It’s really merging three different people with three different sets of expertise into one project,” Garcea said.

In Garcea’s lab, located in the Jean and Jack Thompson Vaccine Research Laboratories of the JSCBB, investigators work on new vaccines such as those for human papillomavirus, a leading cause of cervical cancer that is particularly devastating to women in developing countries.

One corridor away, Randolph’s team, which focuses on creating stable dosage forms for therapeutic proteins and vaccines, developed a process for making vaccines thermostable, or resistant to damage from heat or cold. In this glassy powder state, the vaccine can be stored at temperatures as high as 120 degrees Fahrenheit for three to four months without losing efficacy, Randolph said.

The two began collaborating about two years ago and even formed a spinoff company, Vitravax Inc., which is seeing successful results in vaccine studies conducted in mice.

The Gates Foundation grant will take these innovations a step further by combining the thermostable vaccine powders with techniques developed in the Weimer lab that allow uniform nanoscopic protective layers of aluminum oxide to be applied to vaccine microparticles. This coating process, called atomic layer deposition, not only provides a nanometer-thick protective barrier for the vaccine particles but also helps trigger the body’s immune response.

The trio is now forming extended release, multilayer microparticulate vaccine dosage forms, composed of an inner core of stabilized vaccine coated with aluminum oxide layers and an outer layer of vaccine, all embedded in a glassy powder. When the formulation is injected, the outer layer provides an initial vaccine dose. Next, the aluminum oxide layer slowly dissolves, eventually releasing the inner core which acts as a second dose of vaccine. Patients receive their second or third “dose” without ever knowing it and without a return trip to the doctor.

Although each step of the process has worked independently, researchers cautioned that moving from small test batches in the lab to manufacturing millions of vaccines for public use is a challenging process that may not succeed quickly – or at all.

“We’ve done many of the individual parts of this project,” Randolph said. “Now we’ve got to put those pieces together, and have it work.”

Still, investigators say they’re optimistic about the collaboration, which might never have happened if not for their proximity on CU-�鶹ӰԺ’s East Campus and the interdisciplinary mission of the BioFrontiers Institute, which seeks to drive innovation by combining researchers from different fields.

“One of the hopes (of the BioFrontiers Institute) is that investigators will, by their proximity, do new and interesting things,” said Garcea, who is a member of the Institute. “In a sense, we’ve fulfilled the mission. If the technology works, we’ve really fulfilled the mission.”

The Randolph and Weimer Labs are part of the Department of Chemical and Biological Engineering. The Garcea lab is part of the Department of Molecular, Cellular and Developmental Biology at CU �鶹ӰԺ and the BioFrontiers Institute. At the University of Colorado BioFrontiers Institute, researchers from the life sciences, physical sciences, computer science and engineering are working together to uncover new knowledge at the frontiers of science and partnering with industry to make their discoveries relevant.

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BioFrontiers Faculty Receive BDEG Grants

Anonymous — Thu, 07 Jun 2012 06:00:00 +0000

BioFrontiers Faculty Receive BDEG Grants Anonymous (not verified) Thu, 06/07/2012 - 00:00

BioFrontiers

BioFrontiers faculty receive commercialization grants from the State of Colorado

Ten CU research projects were recently selected to receive grants through Colorado’s Bioscience Discovery Evaluation Grant Program (BDEG-Co). The State of Colorado Office of Economic Development and International Trade began the BDEG program in 2007, providing proof-of-concept grants to move promising CU biotechnologies closer to market readiness, as well as early-stage matching “seed” grants to enable the development and commercial validation of technologies that are licensed from Colorado research institutions by Colorado based start-up companies. Three BioFrontiers faculty members received these awards in the 2011-12 funding cycle:

Christopher Bowman, Department of Chemical and Biological Engineering, CU-�鶹ӰԺ, for inexpensive, highly-efficient synthetic nucleic acids for use in nanoassembly, biodetection and other biofunctional applications.
Heide Ford, Department of Pathology and Department of Obstetrics & Gynecology, and Andrew Thorburn, Department of Pharmacology, CU Anschutz Medical Campus, for a novel biomarker to predict treatment response in solid tumors.
Robert Garcea, BioFrontiers Institute, Department of Molecular, Cellular and Developmental Biology, CU-�鶹ӰԺ, for a next-generation vaccine for human papillomavirus (HPV).
Richard Johnson, Department of Medicine (Renal Diseases & Hypertension), CU Anschutz Medical Campus, for a novel treatment to prevent acute kidney injury following surgery or use of radiocontrast agents.
Malik Kahook, Department of Ophthalmology, CU Anschutz Medical Campus, for an implanted device to reduce intraocular pressure and treat glaucoma.
Uday Kompella, Department of Pharmaceutical Sciences, CU Anschutz Medical Campus, for a new drug to treat “wet” age-related macular degeneration (AMD).
Leslie Leinwand, BioFrontiers Institute, Department of Molecular, Cellular and Developmental Biology, CU-�鶹ӰԺ, for novel drugs to protect from cardiac disease.
David Wagner, Department of Medicine (Pulmonary Sciences & Critical Care Medicine), CU Anschutz Medical Campus, for a drug to prevent/reverse high blood sugar in type-1 diabetes.
Xiao-Jing Wang, Department of Pathology, CU Anschutz Medical Campus, for a drug to treat oral mucositis, a common side effect of radiation therapy.
Hang (Hubert) Yin, BioFrontiers Institute, Department of Chemistry and Biochemistry, CU-�鶹ӰԺ, for more sensitive biomarkers for metastatic cancers and other diseases in body fluids.

“The BDEG award winners this year show an incredible breadth and depth of bioscience research and innovation,” said Tom Cech, Director of CU’s BioFrontiers Institute, an interdisciplinary center designed to explore critical frontiers of unknown biology and translate new knowledge to practical applications. “The BDEG program provides a powerful catalyst to get these ideas out of their academic institutions and into the marketplace.”

The BioFrontiers Institute provided the required matching funds for the grants to �鶹ӰԺ-based researchers Christopher Bowman, Robert Garcea, Leslie Leinwand and Hubert Yin.

�鶹ӰԺ the Technology Transfer Office and the University of Colorado:
The CU Technology Transfer Office (TTO) pursues, protects, packages, and licenses to business the intellectual property generated from research at CU. The TTO provides assistance to faculty, staff, and students, as well as to businesses looking to license or invest in CU technology. Since 2002, 80 companies have been formed based on CU intellectual property; of these, 65 are operational as either stand alone or subsidiary/merged companies.

The University of Colorado is a premier public research university with four campuses: the �鶹ӰԺ, the University of Colorado Colorado Springs, the University of Colorado Denver and the University of Colorado Anschutz Medical Campus. Some 60,000 students are pursuing academic degrees at CU. The National Science Foundation ranks CU seventh among public institutions in federal research expenditures in engineering and science. Academic prestige is marked by the university’s four Nobel laureates, seven MacArthur “genius” Fellows, 18 alumni astronauts and 19 Rhodes Scholars.

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