Posts by JCVI Staff

Scientist Spotlight: Brett Pickett, Ph.D.

The son of a dentist, Brett Pickett grew up in Salt Lake City, Utah focused initially on a career in the family business (his siblings are hygienists and an oral surgeon). Brett believed from an early age that he would follow in his father’s footsteps. He enrolled in Brigham Young University committed to dental school. It was not until Brett’s zoology major was canceled that he became a student of microbiology, where he began researching antibiotic resistance genes in gut microbiota. Dental school was out. Brett received his B.S. in microbiology and continued his studies at the University of Alabama at Birmingham (UAB).

Brett Pickett, Ph.D.

While working in UAB’s bacteria pathogenesis labs, Brett’s path would take another detour as he encountered West Nile, Hepatitis C, and Dengue viruses in his work. He also began to cultivate an interest in computers, technology, and statistics as it related to biological data. These experiences have led to him to his current field of research: viral bioinformatics.

In 2010, Brett moved to the University of Texas Southwestern Medical Center at Dallas to begin his postdoctoral research with Dr. Richard Scheuermann (presently the Director of JCVI La Jolla). Working with Richard, Brett began to shift his focus on how a virus behaves to examining how the human host is responding to being infected. While at UT Southwestern, Brett worked with Richard and his team to identify and develop new statistical, analysis, and visualization tools for the National Institutes of Health (NIH)-funded Viral Pathogen Resource Bioinformatics Database (ViPR). In 2012, Brett moved his family to La Jolla to be a part of JCVI’s informatics team. During this time, his work focused on enhancing the Virus Pathogen Resource and Influenza Research Database bioinformatics resource centers.

Brett stepped away from JCVI for a brief period to work at Thomson Reuters. There he analyzed “-omics” data with pathway analysis and network-building tools, together with drugs and protein target information to better understand viral infection, differences between pathogenic and commensal bacteria, oncology, and other therapeutic areas. This experience allowed him to gain a better understanding of human genetics, disease profiling, and biomarker identification before returning to research at JCVI in 2016.

At JCVI, Brett continues to work on cutting-edge science. He appreciates “the access to collaborators to solve big problems,” and Brett’s efforts are addressing the world’s biggest health challenges. He recently received funding from the US Agency for International Development (USAID) to develop a method for differentiating antibodies against Zika and other closely-related viruses in human patients.

Brett lives in San Diego with his wife and five children. When he is not in the lab, Brett enjoys golf, waterskiing, playing the piano, and visiting the beach with his family. His children, ranging in ages from 1-11, want to be scientists or doctors when they grow up. While there may be no dentists in this generation either, it is clear Brett’s children will have inspirational and accomplished footsteps in which to follow.

Gene Drives Away Mosquitos

For well over a decade, JCVI’s Policy Group has been helping decision makers understand and anticipate the impact of synthetic biology, and where appropriate, devise policies to enhance positive and avoid negative societal outcomes.  Of late, Bob Friedman, who leads JCVI’s Policy Group, has focused on “gene drives,” a powerful new technology that applies gene editing technologies such as CRISPR to quickly “drive” desired traits throughout a population of insects. In 2016, JCVI prepared a report with specific suggestions for the scientific community, US regulators, and international bodies to help advance this promising approach for combatting insect borne human disease and insect agricultural pests, while ensuring that environmental safety and societal issues are addressed.

When University of California San Diego established the new Tata Institute for Genetics and Society (TIGS) with a generous grant from the Tata Trusts, it made perfect sense for TIGS to join forces with JCVI’s policy expertise.  TIGS is starting research centers in both San Diego and India with the goal of engineering mosquitos that can no longer transmit the parasite that causes malaria and, in addition, will rapidly spread that trait throughout a population of mosquitos in the wild.  Friedman’s role is to help both US and Indian regulators who oversee such biotechnologies, as well as relevant international bodies, understand, evaluate, and establish appropriate governance for this innovative technology.  Friedman also serves as a member of a Technical Experts’ Group convened by the United Nations Convention on Biological Diversity to consider oversight of gene drives and other synthetic biology techniques at the international level.

The Leonardo Project

The Leonardo Project has two major components.  JCVI scientists, Karen E. Nelson and Manolito Torralba, aim to use genome sequencing approaches to confirm the identity of the alleged remains of Leonardo da Vinci in addition to characterizing the microbial communities associated with ancient artwork using high throughput sequencing.

Portrait of a Man in Red Chalk (c. 1510) located in the Biblioteca Reale, Turin is thought by many to be a self-portrait drawn by Leonardo da Vinci.

Through collaborative effort with the University of Florence, we collected several sample swabs from a private art collection in December of 2015 in the Tuscany region of Italy.  Sterile swabs were used to collect microbial biomass on each piece of artwork.  Sample artwork included various paintings on wood and canvas as well as marble and stone sculptures.  We extracted DNA from the sample swabs and used 16S PCR to amplify the V4 region of the 16S rRNA gene.  Amplicons were sequenced on Illumina MiSEQ and resulting sequence data was analyzed using a software program called mothur.  Results indicated the presence of known oxidase positive bacteria on samples with paint (canvas and wood) when compared to unpainted wood.  These findings are significant in that oxidase positive genera are capable of metabolic processes that result in water or hydrogen peroxide as a byproduct.  Such byproducts can influence the development of mold.  Furthermore, some oxidase positive genera have demonstrated the capability to metabolize compounds rich in hydrocarbons commonly found in oil based paints.  Finally, we determined that stone/marble and painted samples had a higher level of diversity when compared to unpainted wood.  Our results have provided us with important insight to expand our study further, focusing on other genomics approaches to fully characterize the microbial mechanisms that are influencing the degradation of aging artwork.

The next phase of our study involves sampling additional artwork to confirm and expand on our preliminary findings.  We are targeting artwork that has been designated by art restoration experts as in a state of decay.  Paintings on various substrates; canvas, wood, stone, marble and animal hides are of great interest in addition to frescos and stone, marble, and metallic sculptures. We will generate 16S sequence data along with metagenomics and transcriptomics data.  The metagenomics and transcriptomics data will confirm our hypothesis that various metabolic pathways that are attributed to oxidase enzyme activity correlate with decay of artwork.

We will also collect samples from the remains that are purported to be that of Leonardo da Vinci currently stored in the Saint Hubert chapel in the Chateau d’Amboise.  We will use whole genome sequencing to confirm if the remains are indeed those of da Vinci.  JCVI Adjunct Scientist and Human Longevity Inc., Chief Scientific Officer Dr. Amalio Telenti will be leading a parallel effort to reconstruct the genome of da Vinci from descendants of his relatives who still reside in Florence.

To date we have had several collaborative meetings in Florence, Italy; including three meetings in December 2015, April 2016 and May 2017.  These meetings allowed all participating institutions to present project progress in addition to bolstering collaborative efforts with the varying participants.

Crème de la Crème for Type 1 Diabetes

An effort to engineer insulin-producing microdevice from harmless bacteria of the natural skin microbiotas is beginning

Challenge

Type 1 diabetes (T1D) affects over 1.2 million Americans including 200,000 youth. It is an autoimmune disease where the immune system attacks the insulin-producing beta cells of the pancreas. Beta cells are the central regulator of blood glucose levels in the body. In healthy individuals, beta cells detect blood glucose levels and secrete insulin when the glucose levels rise. Insulin instructs various cells in the body to take up glucose as energy. Maintaining proper blood glucose levels is essential for life in human.

For survival, T1D patients currently rely on insulin injections. Insulin delivery using infusion pumps can be effective, but the current systems require substantial user involvement. They also do not protect T1D patients as well as beta cells. When too little insulin is provided, glucose remains in the blood. The cells then burn fat rather than glucose for energy and produce dangerous levels of ketones as byproducts. When too much insulin is provided, a low level of glucose in the blood also results in a number of problems in the body. There is a limit to how closely T1D patients can monitor and respond to the dynamic changes of blood glucose levels that are also affected by variability in diet and physical activity. A fully automated system with a capacity to adjust insulin dosing based on glucose levels is desirable for managing glucose levels in T1D patients.

Current

Future

Infusion pump

Pancreatic transplant

Infusion pump

Reprogrammed or engineered stem cell

JCVI microbial cream

Closed-loop (automated for glucose level-dependent insulin dosing)

No

Yes

Yes

Yes

Yes

Reliable

Yes/No

Yes/No*

Yes

Yes/No*

Yes*

Non-immune reactive

Yes

No

Yes

Yes/No

Yes

Free from ethical concern

Yes

Yes/No

Yes

Yes/No

Yes

Non-invasive for operation

No

Yes

No

Yes

Yes

Non-invasive for installation

No

No

No

No

Yes

Table. Current and future technologies for T1D therapies. Infusion pumps will soon be automated to allow insulin delivery according to glucose levels. However, from a patient perspective, they are still bulky, labor-intensive, costly, and invasive devices. Pancreatic islet and whole pancreas transplantation has the critical problem of donor shortage. Transplants require immunosuppression with many side effects attributed to the immunosuppressant medicines. Non-pancreatic cells can be modified in the laboratory to produce insulin and then implanted into patients. Cells that are completely reprogrammed to be beta cells would be attacked by patients’ immune systems. Therefore, clever engineering is required. Insulin-producing cells made from bacterial cells that are naturally present in a deep-skin environment, on the other hand, would not be affected by the immune system. They would not have any ethical concern. Moreover, these bacteria can enter the skin without any operation or puncturing of the skin. Regarding the asterisk (*), human systems are much more complex than bacterial systems. Approaches based on human cells would suffer from the lack of precise understanding of systems.

Solution

JCVI is at the forefront of advances in synthetic biology. JCVI researchers have developed powerful and versatile methods for designing and engineering genomes that can be applied to various bacterial species. The solution JCVI is devising for T1D is to give bacterial cells that naturally live in our body the capacity to function like our insulin cells, to produce insulin when blood glucose levels are too high to maintain proper glucose levels in T1D patients. An important ingredient of this study is a set of bacteria recently discovered by our collaborator Richard Gallo at UC San Diego that naturally reside deep within the skin, in layers including the one used for administering insulin in current therapies. These bacteria do not cause problems in our body. Unlike transplants, they do not trigger host immune response. Unlike insulin pumps, they can enter this deep layer of the skin without puncturing the skin.

With funding from Diabetes Research Connection, a pilot project on this idea is underway. To make the deep-skin bacteria function like beta cells, the JCVI research team will introduce a gene into the bacteria for making a version of insulin suitable for production in bacteria. They will also introduce a DNA piece containing three genes for making a glucose sensor to control insulin production in the bacteria. Importantly, they will delete a gene required for making thymidine, an ingredient for DNA, so that their bacteria will be able to grow only when thymidine is provided in the designated hosts within the laboratory. The critical test for this project will be to paint the body of a mouse with their bacteria and see if blood glucose levels drop.

Many rounds of experimentation and improvement will be needed in the future for establishing a system that can be tested in actual T1D patients, but a positive result in this project is expected to produce the excitement in the field to fuel the future efforts toward a bacterial treatment of T1D that circumvents all the struggle of today’s therapies. Support this research!

Figure. Testing the bacterial cream approach in mouse. Bacteria have been found in deep layers of the skin suitable for glucose sensing and insulin administration. The JCVI team will engineer the capacity in these bacteria to turn on an insulin gene in response to high glucose levels. They will also make the engineered bacterial cells dependent on thymidine externally supplied to contain the engineered bacteria within the laboratory and within the designated hosts. The engineered bacteria will be tested in mouse to see if glucose levels can be altered in the treated mice.

Dr. John Glass

Dr. John Glass is a Professor and head of the JCVI Synthetic Biology Group. His expertise is in molecular biology, microbial pathogene­sis, RNA virology, and microbial genomics. Glass is part of the JCVI team that created a synthetic bacterial cell. In reaching this milestone, the JCVI scientists developed the fundamental techniques of the new field of synthetic genomics including genome transplantation and genome as­sembly. Glass was also leader of the JCVI project for rapidly making synthetic influenza virus vaccine strains in collaboration with Novartis Vaccines and Diagnostics, Inc. and Synthetic Genomics, Inc. Glass and his JCVI colleagues are now using synthetic biology and synthetic genomics approaches developed at the JCVI to create cells and organelles with redesigned genomes to make microbes that can produce biofuels, pharmaceuticals, and industrially valuable molecules. Prior to joining the JCVI, Glass spent five years in the Infectious Diseases Research Division of the pharmaceutical company Eli Lilly. There he directed a Hepatitis C virology group and a microbial genomics group (1998-2003).

Dr. Yo Suzuki

Dr. Yo Suzuki is an Assistant Professor in the JCVI Synthetic Biology. His expertise is in genetics, molecular biology, and functional genomics. Suzuki contributed to the JCVI effort to create a minimal bacterial cell. He developed several methods for genome engineering and manipulation during the process. Suzuki also led a project funded by the US Department of Energy for rapidly incorporating synergy among cellulases into a process for breaking down biomass for biofuel applications. Prior to joining the JCVI, Suzuki was a Postdoctoral Fellow at Harvard Medical School studying synergistic genetic interactions among multiple genes in various biological processes (2006-2010).

Summer 2016 Intern Program

Interns in both Rockville, MD and La Jolla, CA participated in our summer 2016 internship program at the J. Craig Venter Institute (JCVI). A total of 19 interns were hired for the summer 2016 program, selected from 578 applicants. Of the 19 interns, six interns were part of the Genomic Scholar Program (GSP) that is a transition program focusing on the leap from a community college to a four-year college using a combination of activities including undergraduate research experience with mentoring and professional development. The interns were mentored by JCVI faculty and research scientists. Mentors design a research project for each intern depending on their education and prior research experience.

GSP interns Emily Samuels, Rolande Tra Lou, Erica Ngouajio, Raja Venkatappa (mentor), Claudia Najera, Kat Rocha, Tayah Bolt (from La Jolla) and Kenya Platero gather at JCVI Rockville's poster session.

GSP interns Emily Samuels, Rolande Tra Lou, Erica Ngouajio, Raja Venkatappa (mentor), Claudia Najera, Kat Rocha, Tayah Bolt (from La Jolla) and Kenya Platero gather at JCVI Rockville’s poster session.

The involvement of fellows in individually focused research projects was designed to stimulate interest in biomedical research as well as to develop independent critical thinking and communication skills with other team members. In addition to research activities, throughout the summer interns participated in professional development activities that included:  education on the importance of documenting research activities and maintaining accurate laboratory records,  responsible conduct of research, the art of reading scientific literature (interns participated in weekly science journal clubs that aimed to teach how to dissect and interpret scientific literature), and scientific presentations. All the interns participated in JCVI internal presentations and presented their summer research as a poster.

Summer 2016 Interns gather for a picture in the courtyard at JCVI La Jolla.

Summer 2016 Interns gather for a picture in the courtyard at JCVI La Jolla.

A brief summary of 2016 interns summer research projects and their mentors are listed below.

Intern Name(s) Research Project Mentor(s)
Roshni Bhattachara Structural implications of unique substitutions found in a paralysis-associated enterovirus D68 clade Richard Scheuermann
Christopher Henderson Assessment of the Contribution of Ascertainment Cohort to the Genetic Architecture of Alzheimer’s Disease Nicholas Schork
Nathan Lian and Anthony Kang Experimental Validation of ChIP-Seq Identified Centremeres Philip Weyman
Rohith Kodukula The Oral Microbiome of Caries in Children: A Study on Twins Andres Gomez
Ian Lamb Identifying Bacterial Antibiotic Resistance Markers in gut microbes Manolito Torralba
Stephanie Mountain Hydrogen peroxide tolerance variation among different isolates of Acinetobacter baumannli Mark Adams and Meredith Wright
Kathryn O’Nell Cell-Type Clustering in Cortical Brain Cells via differential Expression Analysis of Single Nuclei Richard Scheuermann
Josefa Rivera Identifying new promoter elements in Phaeodactylum tricornutum Vincent Bielinski, Philip Weyman, and Chris Dupont
Jennifer Tuman Pan-Cancer Analysis of Somatic Mutations in DNA Damage Repair Genes Alexandra Buckley  and Nicholas Schork
Ben Grimes Using Synthetic Biology Methods to Engineer Herpes Simples Virus-1 and Mycoplasma mycoides subspecies capri Genomes Suchismita Chandran and Sanjay Vashee
Nicolette Maragh Technical Improvements of Sample Preparation for Proteome Analysis Yanbao Yu
Claudia Najera (GSP fellow) Using Synthetic Biology to Engineer Herpes Simplex Virus Type 1 Lauren Oldfield and Sanjay Vashee
Erica Ngouajio (GSP fellow) Developing a method to optimize sequencing of the Zika virus genomic termini Kari Dilley and Reed Shabman
Tayah Bolt (GSP fellow) Reduction of GUS Activity in Phaeodactylum via Episomal hpRNA Expression Philip Weyman
Alexandra Rocha (GSP fellow) Fibronectin and LRG1 protein interactions in T1D patients Rajagopala Venkatappa
Emily Samuels (GSP fellow) Cloning and Expression of proteins in Zika Virus and Legionella pneumophila in E. coli Keehwan Kwon
Rolande Tra Lou (GSP fellow) Filovirus-human protein-protein interaction Reed Shabman and Rajagopala Venkatappa
Carolina Hatanpaa Constructing a Novel Hidden Markov Model for a tRNA Binding Domain Architecture in the Minimal Cell Granger Sutton and David Haft

Scientist Spotlight: Anna Edlund, Ph.D.

Although Sweden is synonymous with Ikea, Volvo, meatballs and ABBA, the country has had a significant impact on science and discovery as far back as the 17th Century. Scientist Anna Edlund, Ph.D. who recently joined JCVI is another Swede pushing the boundaries of discovery in her new role as Assistant Professor, Department of Genomic Medicine.

Anna Edlund, Ph.D.

Anna Edlund, Ph.D.

Anna grew up in the middle of nature on a horse farm in the northern part of Sweden. Inspired by her country’s natural beauty and wilderness, she grew to care a great deal about the environment. During her first years at Södertörn University College she studied ‘green ecology’ and population genetics while she kept her job as a ranger for the Swedish Environmental Protection Agency working in a National park. Dr. Janet K. Jansson first introduced Anna to microbiology during an undergraduate course, and she immediately became fascinated with the unexplored world of microbes – she could not resist becoming a microbiologist. Anna finished her studies at the Karolinska Institute with a Master’s in microbiology and molecular biology. Under the guidance of Dr. Jansson, she pursued her Ph.D. studies in microbiology at the Swedish University of Agricultural Science in Uppsala. Between 2002 and 2007, she studied marine biology specifically exploring the microbial life in sediments of the Baltic Sea. She continued her education in marine microbial ecology as a Postdoctoral Scholar at Scripps Institute of Oceanography at the Center for Marine Biotechnology and Biomedicine, and ultimately returned to Sweden as an Assistant Professor at the Department of Systems Ecology at Stockholm University.

Anna’s trajectory changed in March 2012 when she returned to California at the invitation of Dr. Jeff McLean, a former JCVI scientist and pioneer in the human oral microbiome. As a Project Scientist and Postdoctoral Fellow at UCLA’s School of Dentistry and JCVI, Anna turned her focus from studying bacterial ecological functions in the marine environment towards understanding the role of the oral microbiome in human health.

As a scientist at JCVI, Anna’s research focuses on the complex human oral microbiome and how bacterial gene expression and signaling molecules orchestrate the development of both health and disease associated communities. Anna joined the team at JCVI to work with world-leading experts in microbiology in an environment where most of her time can be spent doing research.

Recently, Anna received a three-year award of $750,000 from the National Institute of Dental and Craniofacial Research (NIDCR) to investigate oral pathogen virulence within complex oral biofilm communities. Her goal is to deepen our knowledge of the molecular processes of oral biofilms during stress and disease-like conditions (e.g. pathogen invasion, low pH). She hopes her findings will lead to improvements in treating and preventing oral diseases.

Research Impact: Accelerating Efforts to Contain and Prevent the Zika Virus (ZIKV)

The rapidly developing Zika virus (ZIKV) outbreak has research groups, government agencies, and industry is all striving to develop a response plan to contain and ultimately prevent ZIKV spread. Currently JCVI is working with both private and public sector funders to sequence and analyze historical and current ZIKV strains. Work at JCVI is geared toward developing sensitive ZIKV diagnostics, significantly increasing the number of ZIKV genomic sequences available, and performing cutting-edge analysis on current and future sequence data.  We expect these efforts to guide the rational design of ZIKV antivirals and vaccines to treat and prevent ZIKV-induced disease.   Here we highlight areas of ongoing ZIKV related work at JCVI.  In each area, additional funding would accelerate our efforts to understand and ultimately control ZIKV infection in the human population.

ZIKV

As of October 12, 2016 the Centers for Disease Control and Prevention reported 3,936 cases of ZIKV infection, with two Florida cities identified as the Ground Zero for local transmission.

ZIKV sequencing efforts at JCVI: 

  • JCVI, through an existing NIH funded grant, is working with the Biodefense and Emerging Infections Research Resources Repository (BEI Resources) to provide high quality sequence data for publically available ZIKV strains. These strains represent a collection of ZIKV isolates, ranging from the initial 1947 isolate from Uganda to 2015 isolates from Puerto Rico, Colombia, and Panama, Mexico, and Honduras. JCVI is providing the gold standard annotated reference sequence for all strains available from BEI and will continue this effort as BEI obtains additional ZIKV isolates.
  • Shortly after the recent Zika virus outbreak emerged in eastern Brazil, Dr. Richard Scheuermann and his bioinformatics team at JCVI collaborated with software engineers at Northrop Grumman to develop a custom Zika website portal to provide genomic sequence and other data about Zika virus through the public Virus Pathogen Resource (ViPR). As of September 2016, the ViPR Zika portal contains 389 genomic and 2399 protein sequences representing the three major Zika lineages – East African, West African and Asian. To support comparative genomics analysis to investigate the evolution of virulence in the newly emerging outbreak isolates, Scheuermann’s group developed an algorithm for predicting the proteolytic cleavage sites that generate Zika mature peptides, and applied this method to produce a comprehensive record of all predicted mature peptides for all Zika genomic sequence in the ViPR database.
  • JCVI is currently working with collaborators in Colombia and Nicaragua to collect sera from patients suspected to harbor ZIKV and to sequence the viral genome from these patients.
  • JCVI was recently awarded NIH supplemental funding to work with Sanofi-Pasteur to screen and sequence human samples suspected to be positive for ZIKV. The majority of samples, provided by Sanofi, are from children and adolescents from the Americas and the South Pacific where mosquito transmitted viruses are common. Over the upcoming year, JCVI anticipates screening both retrospective and prospective human serum samples for ZIKV, with the assumption that many of these samples are from individuals infected with other viral diseases (e.g. Dengue Virus).

Toward the development of a rapid ZIKV diagnostic:

  • Brett Pickett recently received funding from the US Agency for International Development (USAID) to develop a method for detecting antibodies against Zika virus in human patients. A bioinformatics analysis performed previously at JCVI uncovered regions of Flavivirus proteins that differentiate between 10 species of viruses—including Zika. Custom peptide arrays will be constructed to identify immunodominant epitopes in human serum, which we will then optimize as an ELISA-based diagnostic for use in developing countries.
  • To distinguish between ZIKV and other viral diseases, we are developing a highly sensitive and specific ZIKV diagnostic PCR assay.
  • Our assay is sensitive, and we have demonstrated the ability for the assay to identify ZIKV from diverse geographical regions. Future work seeks to move this technology from the laboratory to the field.

Next generation vaccine technology at JCVI can be applied to ZIKV:

  • JCVI has previously coupled synthetic biology for the rapid generation of an Influenza vaccine.
  • Currently, JCVI is using both synthetic biology and vaccinology to develop a universal vaccine for the common cold in partnership with Synthetic Genomics (A company founded by Dr. Venter) and private funders
  • The established vaccine technology at JCVI and our ability to rapidly identify and sequence ZIKV would allow the institute to pursue novel ZIKV vaccine platforms.

Mosquito genomic sequencing:

  • JCVI is currently sequencing the genome of a mosquito that is known to harbor ZIKV and is present in the Americas.
  • Determining the genomic sequence of this mosquito will help research groups identify develop targeted approaches to impair ZIKV replication in the mosquito host.

Current efforts to combat Zika virus involve CLIA-approved methods to detect viral genetic material. In addition, there are multiple players currently developing a vaccine including GlaxoSmithKline, Sanofi, and Onovio Pharmaceuticals. Ensuring that any vaccine doesn’t cause any neurodevelopmental problems further complicates these efforts. Vector control departments around the United States are currently spraying to eradicate adult and larval mosquitoes. While these endeavors serve to prevent virus infection and spread through mosquitoes, they have negatively affected bee populations and organic crops—potentially increasing public acceptance of sterile GMO mosquitoes.

One of the key questions that arose as a result of the Zika outbreak in the western hemisphere is if the virus has mutated to become more virulent, causing more severe neurological pathology than previously circulating strains.  Comparative genomics analysis using sequences and analysis tools in ViPR has identified both nucleotide and amino acid substitutions in the outbreak lineage that warrant further investigation to determine if they are responsible for the apparent increased virulence of the new outbreak strain.  With the detection of mosquito-borne transmission in Puerto Rica and the continental US, there is now a critical need for more funding for further research into the genomic determinants of virulence and for accelerated development of targeted diagnostics, therapeutics and vaccines. Donate today!

Genomic Workshop for Native American College students

A Genomic Science Workshop was held  last week (May 24-26, 2016) at the J Craig Venter Institute Rockville campus for a group of ten Native American college students.  The students participated in two full-day intensive training activities learning how to study the “microbiome” of natural water sources. Each student had the chance to perform hands-on lab work including DNA isolation from an environmental water source, PCR of the 16S ribosomal RNA gene, and gel electrophoresis. Individual computer workstations were provided for the computer lab sessions for students to follow along.  The group was introduced to basic Linux command-line analysis and the popular 16S analysis package QIIME. Overall, the workshop provided the students a foundation of knowledge and tools to identify and classify microbial populations in environmental water sources, and enabled the students to participate in water quality analysis and monitoring efforts of their homeland reservations.

Collage from Maize Cell Genomics Workshop for Undergraduates

Collage from Genomic Workshop for Native American College students

The workshop students were welcomed by JCVI President Karen Nelson and Rockville Campus Director Rembert Pieper. Informal discussion panels were also held to provide networking and research career development opportunities with invited guest speakers including Science Education Directors from the Howard Hughes Medical Institute (HHMI), and Native research scholars from the National Institute of Health (NIH). The students were also presented with a preview of the astronaut microbiomes as an application of human microbiome study. Workshop students also had the opportunity to visit the US Capitol and the National Museum of the American Indian.

The workshop was funded by the National Science Foundation through a Maize Cell Genomics grant and was organized by Agnes Chan (JCVI; co-PI), in collaboration with the Cold Spring Harbor Laboratory (CSHL; PI Dave Jackson, Outreach Educator Joslynn Lee), the University of Wyoming (UW; co-PI Ann Sylvester), Montana State University (Mari Eggers), and the Little Big Horn College (LBHC; John Doyle).  LBHC is a tribal college located in the Crow Reservation, MT. The NSF Maize project has established a long-term outreach relationship with LBHC, and has organized a number of training workshops for Native students previously at LBHC, UW, and CSHL. Participants for the 2016 workshop included Native students recruited from the LBHC, Montana State University, Fort Lewis College, and Northern Arizona State University.

The PIs of the Maize Genomics Project would like to express sincere thanks to instructors from JCVI including Hernan Lorenzi, Yongwook Choi, Vivek Krishnakumar, Stephanie Mounaud,  the JCVI Information Technology team, the Administrative Assistant team, and all colleagues for their generous assistance, support, and patience for a successful outreach educational workshop.

To find out more information on workshop schedule, notes, and manuals, please visit the Maize Cell Genomics project web site at http://maize.jcvi.org/cellgenomics/2016_pcr.php.

Ongoing Zika virus work at JCVI

The rapidly developing Zika virus (ZIKV) outbreak has research groups, government agencies, and industry all striving to develop a response plan to contain and ultimately prevent ZIKV spread. Currently JCVI is working with both private and public sector funders to sequence and analyze historical and current ZIKV strains. Work at JCVI is geared toward developing sensitive ZIKV diagnostics, significantly increasing the number of ZIKV genomic sequences available, and performing cutting-edge analysis on current and future sequence data. We expect these efforts to guide the rational design of ZIKV antivirals and vaccines to treat and prevent ZIKV-induced disease. Here we highlight two areas of ongoing ZIKV related work at JCVI.

Zika virus

This is a digitally-colorized transmission electron micrograph (TEM) of Zika virus, which is a member of the family Flaviviridae. Virus particles, here colored red, are 40 nm in diameter, with an outer envelope, and an inner dense core. Image credit: CDC/ Cynthia Goldsmith

JCVI/BEI Resources/NIAID: JCVI, through an existing NIH funded grant, is working with the Biodefense and Emerging Infections Research Resources Repository (BEI Resources) to provide high quality sequence data for publically available ZIKV strains. These strains represent a collection of ZIKV isolates, ranging from the initial 1947 isolate from Uganda to 2015 isolates from Puerto Rico, Colombia, and Panama. JCVI is providing the gold-standard annotated reference sequence for all strains available from BEI and will continue this effort as BEI obtains additional ZIKV isolates. A list of the ZIKV isolates sequenced by JCVI are found here: https://www.beiresources.org/Organism/118/Zika-virus.aspx

JCVI/Sanofi-Pasteur/NIAID: JCVI was recently awarded NIH supplemental funding to work with Sanofi-Pasteur to screen and sequence human samples suspected to be positive for ZIKV. The majority of samples, provided by Sanofi, are from children and adolescents from the Americas and the South Pacific where mosquito transmitted viruses are common. Over the upcoming year, JCVI anticipates screening both retrospective and prospective human serum samples for ZIKV, with the assumption that many of these samples are from individuals infected with other viral diseases (e.g. Dengue Virus). To distinguish between ZIKV and other viral diseases, we are developing a highly sensitive and specific ZIKV diagnostic assay. After confirming ZIKV positive samples, JCVI will perform whole genome sequencing and sequence analysis to understand the evolution of the virus over time and geographical location. We hope that results from this collaborative work will significantly increase our understanding of the origins of the ZIKV outbreak in the Americas and lay the groundwork for future collaborations with NIAID and Sanofi.

Unlocking the Mysteries of the Microbiome

In the early 2000s, JCVI researchers pioneered in the exploration of the human microbiome, the community of microbes that live in and on the human body. Originally while at The Institute for Genomic Research (TIGR, now part of JCVI) Drs. Craig Venter and Hamilton Smith were awarded a grant from DARPA to examine the microbes found in the human gut.  This work was carried out by researchers at JCVI and published in 2006 in Science.  While this team had previously published 16S surveys of the human body, this paper in which the researchers found more than 60,000 microbial genes was the first metagenomic description of microbes resided anywhere on the human body.  Ten years since this seminal publication, our scientists continue to pave the way for a broader understanding of these vast microbial populations.

fragment recruitment plot

Visualization of ocean microbial data collected on JCVI’s Global Ocean Sampling Expedition (GOS). The Sorcerer II circumnavigated the globe for more than two years, covering a staggering 32,000 nautical miles, visiting 23 different countries and island groups on four continents.

On May 13, 2016, Drs. Craig Venter and Karen Nelson were present at the White House for the launch of the National Microbiome Initiative (NMI).  The NMI will invest $121 million in new microbiome studies in fiscal years 2016 and 2017.  The goals of the project are to supplement fundamental research, develop new technologies and engage more people in this area of research.

Today we know that the human body is host to more than 1 trillion microbes. Thanks to continued advances in genome sequencing technologies and metagenomic analysis JCVI scientists are providing a deeper understanding of these microbes across a variety of fields. JCVI researchers know that translating the role of the microbiome in the development of health and disease in humans is essential.  We believe that eventually the screening of the human microbiome will be a routine part of medical care, leading to prescribed diets and preventive measures personalized to an individual.

JCVI currently has several dozen microbiome studies underway.  In this issue of Amplifier, we are highlighting some of our most exciting and cutting edge work unlocking the mysteries of the human microbiome.

The Effects of Long-Term Space Travel on the Microbiome of Astronauts

On March 1, the world celebrated the safe return of NASA astronaut Scott Kelly after 340 days in space.  Researchers are fascinated to learn more about the impact of long-term space travel on the human body, and JCVI scientists are excited to be a part of the process.  During a mission to space, astronauts are subject to many stressful conditions (g-forces, radiation, microgravity, anxiety, etc.) that can have a negative impact on their health. For example, astronauts lose muscle mass, bone density, and experience a wide range of health problems with everything from their vision to their gastrointestinal tract. Several studies have demonstrated that space travel also affects the astronauts’ immune systems (for example the reactivation of latent viruses like Herpes Simplex Virus 1 and Epstein Bar virus) and have shown some evidence suggesting that stool microbes change after space flight.

JCVI researchers want to determine how the composition of the astronauts’ microbiome changes during long-term space missions (six or more months), and to evaluate potential risks to astronaut health from changes in the microbiome. We are also interested in how the microbiome of astronauts interacts with other factors such as the microbial communities that inhabit the International Space Station (ISS). To accomplish this, we will monitor the astronauts’ health status, environmental stress, and exposure to space conditions. The skin, tongue, nose and gut of each astronaut will be sampled at multiple time points before, during, and after the mission to the ISS. By sampling the microbiome of astronauts on earth while in peak physical health and during subsequent space flight, we will be able to define signatures of human response to a variety of relevant aspects of space travel. Astronauts will also sample different surfaces and the water supply during their stay at the ISS to correlate crew microbiomes with the microbes living at the ISS. We will also assess changes in the astronauts’ immune function and stress levels throughout the mission by analyzing their saliva and blood for metabolic markers. Finally, we will correlate the microbiome and immune function data collected with other measured metadata including astronaut health and hygiene as well as environmental factors such as temperature, humidity and environmental factors.

This research program is being led by Dr. Hernan Lorenzi.

The Gut Microbiome and Human Evolution

Who we are, where we come from and how we came to be as we are, are questions that have always fascinated biologists. The reasons to answer these questions are multiple, but one critical aspect centers on understanding what makes us human. To start addressing these issues JCVI scientists are exploring the gut microbiome of non-human primates, our closest living relatives, and of populations that most faithfully reflect the lifestyles of early hominids: hunter-gatherers. The goal of this project is to establish an evolutionary baseline to shed light on the host-microbe factors that impacted health and disease in modern and western human populations.

Our scientists have shown, in several recent publications, that the gut microbiome of wild gorillas, is strongly shaped by the external environment, namely by diet. Specifically, we showed that gut microbes adapt to different dietary stimuli, probably providing gorillas with energetic plasticity when preferred feeding resources are seasonally and temporally absent. Interestingly, we also suggested that, in conditions in which gorillas exploit high-energy diets, their gut microbiomes resembles those of humans. This fact has critical implications to understanding the evolutionary origins of obesity and inflammation in modern human populations from a microbe perspective. Along these lines, our most recent publication on the gut microbiome of central African hunter-gatherers, traditional agriculturalists and western humans shows evidence that transitions to agriculture and industrialization, and giving up hunting and gathering could have radically changed our gut microbiomes for good. This observation is vital considering that traditional hunter-gatherers, whose microbiomes resemble those of wild gorillas, do not show symptoms of modern inflammatory disease. These observations highlight the potential impact of gut microbes in human evolution.

The research team consists of  Drs. Andres Gomez and Karen Nelson.

Solving Crimes with Your Microbial Signature

In January 2016, JCVI received a two-year, $962,500 award from the United States Department of Justice to design and build an open-access microbiome database for the forensic science community. The Forensic Microbiome Database (FMD), the first of its kind will be populated with several thousand microbiome datasets and associated metadata available from the public domain. The database will be based on established procedures for database development designed at the JCVI, incorporating expansive sets of data and metadata that relate to forensic evidence.

The goals of this project are to: provide a host location and continuous monitoring of the database; define well-structured standard operating procedures for data generation and searching against and uploading data into the FMD; and test the utility of the FMD by sequencing a range of samples obtained geographically for querying and proof of concept against the database. The foundation of this project will serve for future enhancements of the FMD and utility for forensic casework. The research team expects this will become the community resource for analysis of microbiome data and for attributing weight to microbial forensic evidence.

The research team consists of Rhonda Roby, Lauren Brinkac, Toby Clarke, Andres Gomez, Karen Nelson, Harinder Singh, and Shibu Yooseph,

Using the Microbiome to Advance Wound Therapies

Chronic wounds are wounds that fail to heal after 4 months of proper wound care and management.  It is a major public healthcare burden that affects an estimated 1% of the US population and costs $25 billion per year. Common chronic wounds are leg, foot, and pressure ulcers occur in adults especially the elderly with diabetes, vascular diseases, or specific body locations under prolonged pressure. According to the Centers for Disease Control and Prevention, approximately 12% of U.S. adults with diabetes had a history of foot ulcer and 11% of U.S. nursing home residents had pressure ulcers. In addition to the economic burden, from the perspective of patients chronic wounds can also lead to loss of function (e.g. amputation), decreased quality of life, and increased rate of mortality.

At JCVI we are interested in deciphering not only the microbial communities present in chronic wounds but also their potential impacts and relationship with the wound healing outcome, for working towards more effective clinical strategies of wound healing. In collaboration with George Washington University, we are conducting a study to analyze chronic wounds. Samples are selected from patients enrolled in the Wound Etiology and Healing biospecimen and data repository (WE-HEAL). We will analyze the chronic wound microbiome at the molecular level, and attempt to identify biological indicators that can be used to predict the healing outcome to further advance wound therapies and management.

This project is being led by Dr. Agnes Chan.

Metagenomic Epidemiology of Antibiotic Resistance in Infectious Diarrhea

Genes that encode antimicrobial resistance (AMR) to antibiotics have been detected in environmental, insect, human and animal metagenomes and the sum of these are known as “resistomes.” While metagenomic datasets have been mined to characterize the healthy human gut resistome, directed metagenomic sequencing has not been used to examine the spread of AMR. Especially in developing countries where sanitation is poor, diarrhea and enteric pathogens likely serve to disseminate AMR elements of clinical significance. Unregulated use of antibiotics further exacerbates the problem by selection for acquisition of resistance. This is exemplified by recent reports of multiple AMR in Shigella strains in India, in Escherichia coli in India and Pakistan, and in nontyphoidal Salmonella (NTS) in South-East Asia.

Sarah Highlander, Ph.D. and her team are characterizing the microbial composition and its component AMR transfer elements (such as plasmids and transposons) by metagenomic sequencing of stool samples from pediatric patients from Colombia who are suffering from diarrhea. Our goal is to assess whether groups of species/strains associate with specific mobile genetic elements and whether their presence is enhanced or amplified in diarrheal microbiomes. This work could potentially identify clonal complexes with enhanced resistance and potential pathogenesis.

For more information on how you can support or human microbiome research program at JCVI, please contact development@jcvi.org.