Malaria Genome Focus Group Document
On April 7, 1997 the Burroughs Wellcome Fund sponsored a Malaria Genome Focus Group Meeting, which was convened by the Malaria Foundation International in Rockville, Maryland, USA. This document has resulted from the contributions of the participants attending this meeting and the contributions of others in the scientific community who chose to provide feedback on this paper as it was being developed. Additional comments remain welcome at MalariaGenome@malaria.org
Movement between April and October 1997:
- Discussions for the establishment of a global repository(ies) for malaria research material and reagents continue. Initial funding for such a resource is being obligated by the US-NIAID/NIH. Further, a NIAID planning meeting with malaria researchers is scheduled for November 24-25 to obtain advice and set priorities for initiating the development of this resource. More current information is available at www.malaria.org/NIAID.HTM. Suggestions can also be sent to email@example.com.
- Strong interest has been expressed for the development of a very practical user-friendly centralized database for analyzing and utilizing malaria genome sequence information. Discussions continue to determine how best to make this a reality.
- Suggestions have been made for the development of a rodent malaria focus group to discuss species-specific issues.
- A vivax malaria network is being initiated by a core group of researchers who have taken part in the discussions presented in this document.
- Funding has been committed from the NIH/NIAID for obtaining P. vivax and rodent malaria EST/GSTs, and discussions have continued to determine which species/isolates should become the focus.
- Requests continue to be made for suggestions on complementary projects.
For subsequent meeting reports please refer to the MFI's Malaria Genome Database page at www.malaria.org/genome.html
Malaria Genome Focus Group Working Document
Introduction of the current malaria genome project and update on progress
Recognizing the recent advances that have been made in high throughput sequencing of DNA and in bioinformatics and especially their application to sequencing the genomes of bacterial pathogens, a number of investigators considered, in late 1995/early 1996, undertaking the complete sequencing of the genome of Plasmodium falciparum. Given the potential benefits of this project, several funding agencies expressed an interest in providing support and collaborating on this project. A meeting, co-organized by the National Institute of Allergy and Infectious Diseases (NIAID) and the Burroughs Wellcome Fund (BWF), was held in May 1996 to coordinate the activities of the various funding agencies. In addition to NIAID and BWF, representatives from the Department of Defense (DoD) and the Wellcome Trust (WT) participated in the meeting. At that meeting, the various organizations agreed to coordinate their activities and announced the following:
All sequencing groups agreed to work on the 3D7 strain of P. falciparum.
The WT announced that it would support the Sanger Centre with malaria expertise provided by investigators at Oxford University.
The NIAID announced that it would support, with funds from the NIH Office of Research on Minority Health, sequencing efforts at The Institute for Genomic Research (TIGR) in collaboration with Naval Medical Research Institute (NMRI).
The BWF agreed to provide support for pilot projects which would address some of the technical hurdles involved in handling and sequencing P. falciparum DNA.
The DoD representatives agreed to go forward with a proposal to DoD for support of the project.
In December 1996, a meeting, again co-organized by NIAID and the BWF, was held in Baltimore, MD. The meeting brought together representatives from the sequencing centers (NMRI/TIGR and Oxford/Sanger), from the pilot projects and from the funding agencies. In addition, there was participation of investigators involved in the P. falciparum chromosome mapping project and the EST/GST projects.
Efforts have been underway for several years to provide maps of the P. falciparum chromosomes. Status of that project can be found on the Internet at <http://www.wehi.edu.au/biology/malaria/genomeInfo/MapData/MapData.html>. The University of Florida has undertaken the sequencing of Expressed Sequence Tags (ESTs) of cDNAs from blood stage parasites and Genome Sequence Tags (GSTs) of a mung bean nuclease library. The goal of this project is to identify approximately 75% of P. falciparum genes. At present, the project has sequenced approximately 3000 tags. Details of the status of this project can be found on the Internet at <http://parasite.arf.ufl.edu/malaria.html>.
As a result of the discussions and review of progress made, the investigators and funding agencies agreed, at the December 1996 meeting, on the following:
Increased effort should be given to shotgun sequencing of isolated whole chromosomes. Sanger Centre will complete chromosome 3 and TIGR was to initiate sequencing of chromosome 2. Since that meeting, these sequencing centers and a group at Stanford have agreed upon dividing the remaining chromosomes among themselves. Results of initial chromosome 3 shotgun sequencing efforts can be accessed on the Internet at <http://www.sanger.ac.uk/pathogens/>.
The groups involved in the sequencing project should meet again in June 1997 to review progress. A meeting has now been scheduled for June 16-17 at the Sanger Centre. (Update: this meeting took place as scheduled and the Wellcome Trust has prepared a report).
Effort should continue on projects to facilitate the full-scale cloning effort, including generation of large insert libraries, overcoming clonal instability, gap closure, etc.
Discussion should be held within the broader malaria community on additional malaria genome projects that would advance research and would lead to development of diagnostics, drugs and vaccines. Issues under consideration included which other malaria species (P. vivax and/or rodent malaria, other) should be sequenced either as full scale sequencing or more limited EST or GST projects; also, are there other projects which would complement the large scale sequencing effort underway for P. falciparum?
Also under consideration were plans to begin to think ahead about ways to capitalize on the genome information; e.g. comparative genomics, whole genome approaches to functional analysis.
Thought should be given to mechanisms by which the information derived from the genome project would be made available to the entire malaria community in a manner that would promote its optimal utilization.
It is these last three points that formed the impetus for the Malaria Genome Focus Group Meeting and subsequent discussions in the malaria research community.
At the request of the Burroughs Wellcome Fund, the Malaria Foundation convened the Malaria Genome Focus Group, which was held on April 7, 1997 in Rockville, Maryland, USA. This group (participants listed below), came to a general consensus on the following points up for discussion. Additional considerations provided by other members of the scientific community, also listed below, have since been condensed and incorporated as summarized commentaries.
The URL links mentioned above and others can be found at the Malaria Foundations web page designated "For the Scientific Community," <http://www.malaria.org/SCICOMM.HTM>.
POINTS for DISCUSSION
A. How can the sharing of information and resources relevant to malaria genome research be improved?
1. Networking is crucial for the success of the overall malaria genome project.
2. Networking will involve the sharing of information as well as the sharing of resources and will help to involve a larger community of scientists in malaria genome initiatives.
3. The malaria genome project will provide a platform from which new approaches to malaria research can be pursued and public awareness of malaria can be enhanced.
4. The scientific community can pro-actively aid this process, which ultimately could lead to greater resources and productivity towards the development of malaria drugs, diagnostics, and vaccines.
5. Importantly, effective networking will also help to attract investigators from other specific disciplines, such as biochemistry, cell biology, or immunology, to malaria research.
Comments (summarized and condensed)
Networking and the sharing of information and resources are crucial, and the malaria genome discussions should be kept open in the broad scientific community, particularly since the output should reflect the wishes of the research community. There is a strong desire for others to enter the discussions that have begun, and to broaden the forum for debating the management and use of information obtained from the malaria genome sequencing projects. Although networking can be difficult to achieve, especially given everyones time constraints, the importance remains obvious. Cooperative approaches will help to maximize the use of the information with what financial resources are in-hand and without unnecessary overlap. Besides, other genome projects have shown that basic biology can advance rapidly when the information is used by a wide variety of researchers.
It was further stressed that this is a global project. Although the sequencing collaboration described has been largely a US/UK venture, it is important that these efforts are not seen as exclusive. Other groups currently play facilitating roles, and attention on the efficient dissemination and use of this information worldwide was considered crucial. Additionally, the importance of involving scientists from malaria endemic countries was stressed, with special emphasis on their need to be able to electronically access malaria genome information from databases.
B. What are key biological questions that can be addressed once Plasmodium genomic DNA sequence is in-hand?
1. What are the complete biochemical pathways present in malaria parasites? Which of these may be targets for future drug development? What can the biochemical pathways tell us about the biology of these organisms?
2. What parasite gene products modulate host responses, and what are the genetic determinants for virulence and pathogenesis? What parasite gene products are obvious targets for vaccine development (e.g. surface proteins, enzymes etc.)?
3. How are malaria parasite organelles unique, and what are their functions?
4. What can we learn from comparative genomics; i.e. comparison of malaria genome data with genome information of other organisms?
5. What are the mechanisms of Plasmodiums genetic diversity, including antigenic variation, drug resistance, evolutionary diversity, and mutation frequency?
6. How does Plasmodium genetic diversity influence the epidemiology of disease, and how can we more effectively approach this question with new genomic information? How can this information be utilized to develop more specific diagnostic tools or malaria vaccines?
7. What are the determinants of sexual differentiation, stage-specific expression, and differential expression in response to environmental stimuli?
Comments (summarized and condensed)
Overall this was viewed as a thorough listing of key biological questions to be addressed. An additional point made was that the information from a single genome would be only a starting point for understanding the antigenic repertoire for vaccine development. However, once one P. falciparum genome is sequenced, high throughput sequencing will not be required to determine the antigenic repertoire for most vaccine candidates. On the other hand, analysis of the var gene repertoire, if to be accomplished, will require sequencing of large amounts of DNA of the order of 0.5-1.5 Mb per isolate, and would require the involvement of a number of laboratories. On another note, it was mentioned that knowledge of differential expression in different hosts during the life cycle might be instructive.
Several individuals mentioned funding concerns, and it was stressed that basic funding to maintain many productive laboratories is already limited. Several contributors also indicated that while this document is quite comprehensive and well thought out, it lacks emphasis and direction (short and long term) regarding how best to accomplish objectives of utilizing genome information to answer biological questions, especially in a cost-effective manner. Follow-up discussions should address these issues, and it was suggested that a list of action items specifying what is being done by whom, and, perhaps, how and by when, be included in this and subsequent reports on malaria genome projects.
C. Should the genome(s) of other Plasmodium species be sequenced in parallel (i.e., in addition to P. falciparum)? If so, which genome(s)? Also, should the approach for other species be whole genome sequencing or EST/GST sequencing?
1. Additional malaria genome information from other species of Plasmodium would be valuable to the scientific community, both for understanding key biological features of malaria parasites, and for the development of malaria drugs, diagnostics, and vaccines. Moreover, genome information from other malaria species provides the basis for the use of model test systems and an important level of comparative genomics between highly divergent groups of Plasmodium species.
2. The consensus of this group is that the focus of other efforts should be on Plasmodium vivax and on a rodent malaria species.
3. Many participants considered it important for P. vivax to be included in the malaria genome sequencing efforts because 1) it is a major human pathogen, 2) it is evolutionarily distinct from P. falciparum, and 3) unlike P. falciparum, the DNA of P. vivax is of lower overall A/T content and is generally stable in E. coli. Acquisition of P. vivax DNA for genome work is feasible, as are biological studies, particularly within the framework of network support.
4. Genome sequence information from a rodent malaria species was considered important because 1) such a model system can be manipulated and is generally available to large and small laboratory groups, and 2) given the recent successes in genetic transformation of Plasmodium, a rodent model system will be especially important for studying the general biology and genetics of malaria parasites. While no conclusions were reached, the majority of the discussion focused on P. berghei and P. yoelii as candidate rodent genomes, with some inclination towards the former since it can be cultivated in-vitro and has been genetically transformed with success.
5. P. vivax and rodent malaria genome sequencing should be carried out in parallel with the P. falciparum project, and need not compromise the success of this project. It was generally agreed that the P. vivax and rodent malaria genome projects would be greatly facilitated by the development of cost-effective focused research networks, and that, rather than being detrimental to the P. falciparum project, would, in fact, provide valuable information to this project.
6. Pilot malaria genome projects on other malaria species should be initiated and supported, with an initial focus on EST/GST sequencing and chromosome mapping, and not necessarily, at this immediate time, on additional whole genome sequencing projects. The EST/GST approach is both low in cost (estimate is ~$10/EST/GST, based on the costs of the Toxoplasma gondii project) and will produce valuable data in a short time horizon. The minimal target for these pilot projects should be ~15,000 ESTs/GSTs per genome, based on an estimate of 7,500 genes. This compromise consensus, on an initial focus towards EST/GST sequencing, should not preclude whole genome characterization if scientific and fiscal evaluations indicate such an undertaking(s) is feasible and desirable, particularly in the case of P. vivax.
Comments (summarized and condensed)
Much discussion was raised about another genome. Several people emphasized that P. falciparum is the first priority, and that given the current funding situation additional sequencing efforts may be difficult to consider at this point. If another genome project were to begin, support was for the EST/GST sequencing approach, rather than whole genome sequencing. Obtaining ESTs/GSTs is relatively inexpensive, and they provide good "seed data" even though the corresponding sequences are not complete. Although the importance of P. vivax as a human parasite and the value of P. vivax ESTs/GSTs was emphasized, several individuals stressed the greater accessibility of rodent malaria to the broad malaria community (i.e. many more laboratories can work on rodent malaria) and the usefulness of these models to approach important biological questions such as cytoadherence. For these reasons, if entire genome sequencing were to proceed for one additional Plasmodium species, support for a rodent parasite outweighed that for P. vivax. Although rodent malaria models continue to be generally recognized as more amenable to biological studies by a larger number of laboratories, hope was expressed among those working with P. vivax that continued development and support of a P. vivax network would overcome several of the existing difficulties for a number of groups to study P. vivax. Recent advances in culturing P. vivax may also facilitate future studies on this parasite.
There were many comments specifically related to the question of which rodent malaria parasite(s) should become a priority, and there was not a clear consensus. It was noted that, ideally, one should consider the availability of the life cycle, the ability to transfect the parasite, the availability of data on cytoadherence, the availability of antigenically diverse isolates, and whether or not one better represents a biological model for P. falciparum especially with regards to pathogenesis. Several researchers provided support for the consideration of P. chabaudi, with the argument that it provides a more suitable biological model for P. falciparum than the others and provides a model for understanding questions on antigenic variation and sequestration. It was also noted that transfection vectors are available for P. chabaudi, but whether or not P. chabaudi transfection has been achieved was not mentioned. A number of additional characteristics were highlighted "for P. chabaudi," which may be best as part of a full paper on the subject, comparing and contrasting the different models. It was also noted that one priority should be to determine if transfection experiments have been or could be performed with rodent malarias other than P. berghei. P. berghei was supported because it has the advantage of an in-vitro system and has been transfected, but the limitation of an apparent lack of genetically diverse isolates was noted. The question was also posed as to whether other species could be cultured. Other support was expressed for P. yoelii as a model for vaccine development utilizing sporozoite challenge. Some additional discussion has followed on the possibility of obtaining EST/GST information from more than one rodent parasite. It could be extremely useful for researchers working with the different rodent malaria models to meet and elaborate on the pros and cons of each of their systems.
Additional comments were made about the potential need to sequence more than one isolate of P. falciparum, at least for targeted regions of the genome, to understand about sequence diversity especially for particular biological questions such as agglutination or cytoadherence.
D. What tools/reagents will facilitate full utilization of Plasmodium genomic information that is generated? What complementary projects should be initiated now to maximize the use of incoming malaria genome information?
1. Standardization of parasites (i.e. isolates/strains generally used in malaria research laboratories), gene banks, and repositories for selected reagents is required. Malaria research, today and as new sequence information becomes available, would be greatly facilitated by the general availability of high quality and well-characterized genomic DNA and cDNA libraries for both human malaria parasites and selected non-human malaria models. Further, a central reagent repository(ies) could include a variety of tools such as specific monoclonal antibodies and antisera, recombinant proteins, indexed and tagged cloned ESTs and GSTs, YACs, and BACs*, transformation vectors, and standardized parasite strains. Designated facilities for research requiring the use of specialized animal models or insect stages would also provide valuable complementary support if made generally available and accessible.
2. Comments are specifically requested to learn what malaria researchers in the broad scientific community might like to see available in repositories. Please prioritize your suggestions. Also, what special resources or networks would be especially useful to this field?
3. The well-recognized, generally high instability of P. falciparum DNA sequences in E. coli, as well as the presence of frequent very long stretches of A or T sequences, are likely to be significant problems in the derivation of the complete and accurate P. falciparum genomic sequence. Hence, it is recommended that a systematic effort be undertaken to explore various strategies to overcome this problem. These could include identifying alternative host-vector systems, or identifying E. coli mutants in which large fragments of P. falciparum DNA can be stably maintained.
4. One immediate application of sequence information will be in the area of molecular genetic analysis. Several necessary molecular tools now exist, but improvements in their application and efficiency of use are of high priority.
a. Basic genetic manipulation of malaria parasites should be further developed and improved. This will require the identification and utilization of additional selectable markers, the enhancement of transfection efficiency, as well as the identification of promoter/regulatory regions for differential and stage-specific expression.
b. Complementation analysis capabilities should be developed, which could include the use of heterologous systems such as yeast and Toxoplasma gondii, as well as the malaria parasite itself. This will be facilitated by the identification of centromeres for chromosomal complementation and the development of Plasmodium artificial chromosomes.
c. Information and materials from currently recorded laboratory genetic crosses is extremely valuable and should be made generally available to the malaria research community. The development and analysis of additional laboratory genetic crosses, as well as studies aimed at comparatively analyzing natural crosses, should also be given consideration.
d. Possibilities for developing attenuated parasites should be explored.
5. Culture Technology should be expanded to include the further development of:
a. P. falciparum exoerythrocytic culture systems, in-vitro and in-vivo, such as in SCID mice;
b. culture systems for other Plasmodium species and stages of the life cycle; and
c. P. falciparum axenic culturing systems.6. Whole Genome Expression Screening Technologies should be developed for Plasmodium.
a. The latest technologies for utilizing the DNA sequence information of entire genomes to best understand the complex phenotypes of the corresponding organisms should be explored. In particular, the recently developed "DNA chip technology" should be investigated for applications in malaria research.
b. Technological advances in expression screening could also aid efforts to assess differential expression, for example, due to environmental stimuli or var gene switching.
c. Expression screening advances could also have especially important applications in the area of malaria diagnostics
7. Homology Cloning projects could be valuable for obtaining and studying selected genes from other Plasmodium species.
* YACs = Yeast Artificial Chromosome
BACs = Bacterial Artificial Chromosomes
Comments (summarized and condensed)
It was noted that the need to develop means of stably cloning large fragments of P. falciparum is critical for utilization of the genome information, and more resources should be dedicated to solve this problem; stability is initially important for obtaining reliable sequence, but will also be important for conducting biological studies. It was also noted that mapping of EST/GST sequences to YACs would provide valuable landmarks for assembling and ordering sequence. Additionally, the problem of low transfection efficiency was emphasized as a barrier to full utilization of the sequence information; thus, investigations to improve transfection efficiency should be a priority. Complementation of heterologous systems clearly should be exploited, but whether a large effort on P. falciparum artificial chromosomes should be a priority was questioned. Overall, the suggestion for the development of new genetic tools is a good one, but new funding sources will need to be developed to turn this into reality.
It was also mentioned that Yac contigs for chromosomes 3, 4, 5 and 12 are being produced at WEHI and distributed to the Sanger and Stanford groups to facilitate sequencing and closure of these chromosomes. Available data on these contigs should be available at a web page set up by Ross Coppel and, some of it is also included at the Sanger Center web site. In addition, it was noted that sequencing of existing PAC library (RPCI-91 and RPCI-92) inserts (7-8 kb) may help to fill in existing gaps between contigs.
Many comments addressed the establishment of a repository or repositories, how they might be set up, and how they would be funded. It was emphasized that a mechanism(s) for long-term funding was especially critical. In addition to items mentioned in the text above, it was indicated that parasite lines should be standardized by a set of internationally agreed upon criteria, and that the culture history of each line from whichever sentinel stock is used should be known. Well characterized, strictly defined and representative libraries could be useful in some cases for P. falciparum (ideally 3D7), P. vivax and rodent malarias; however, in many cases defined primer pairs and PCR protocols could replace the need for DNA libraries. A desire for YAC libraries, yeast for growing and expanding YAC libraries, plasmids which are used for sequencing, and bacterial strains which people have found good for malaria DNA stability, was also expressed. It was also noted that stage-specific and organelle-specific markers (e.g. specific antibodies) for P. falciparum, P. vivax and rodent malarias would be valuable in a repository. Support for the development of a catalogue/database directory of reagents with their sources and properties was also expressed by several individuals. The issue of corporate rights to certain reagents was also mentioned as a barrier to the availability of some reagents, and that up-front discussions with companies, before products are produced, might enable the development of mechanisms to place future products in repositories.
It was also noted that the assembly and dissemination of reliable protocols would be very valuable especially for scientists beginning to evaluate genomic information in developing countries. Basic protocols, not involving the use of expensive kits, could include standardized methods for the isolation of DNA and RNA from parasitized blood samples, and PCR and rtPCR conditions for analyses involving specific genes or gene regions. These would facilitate the widespread use of the genome information.
E. What is the best way to facilitate access to information generated in malaria genome projects for the broader scientific community? How can the current databases be linked and the information be shared in a common format?
1. It was a general consensus of the meeting that there was an immediate need to facilitate the sharing of available Plasmodium genomic sequence information, and to make these data available and understandable to the widest possible scientific community.
2. A central and coordinated site for Plasmodium genome sequence information should be established and maintained. It is recommended that this site be located in one of the major genome reference centers, such as Genbank, and that a dedicated staff member be employed to facilitate access to and the coordination of malaria genome information.
a. This site should be user friendly and include educational tools for the user, with access to any necessary training at the site.
b. This site should also have a variety of relevant searching options.
c. This site should contain an ORF database, catalogued EST and GST information, chromosomal maps, contig maps, and data from genetic crosses concerning inheritance, among other relevant information.
3. Both finished and unfinished sequence should be collected and made available at this site. Immediate release of sequence information is strongly supported. Notwithstanding, investigators should be made aware of potential pitfalls in working with immediate-release sequence information. Sequence releases should include trace information.
4. The current information from the YAC based mapping of P. falciparum chromosomes should be collated and made generally available to the scientific community as rapidly as possible. Efforts should also be made to start placing available ESTs and GSTs, as well as microsatellite markers, on these maps, also within a short time frame. This data will likely be important for the whole genome sequencing of P. falciparum.
5. It was also suggested that a common interface be developed so that a variety of databases and search engines could be easily interrelated.
Comments (summarized and condensed)
There is clear frustration with the currently available genomic DNA databases, as it is difficult for a biologist/malariologist to readily obtain much useful information from them. Besides, it was noted that simply handing our needs over to Genbank personnel would be a mistake, as they have no particular expertise in genome information as opposed to sequence information and, additionally, have no particular expertise in malaria.
Very strong support was expressed for the development of a database that would allow easy access to information and would allow biological questions to be approached directly (and without the need to rely on the aid of a bioinformatician, which for many groups is not even available on-site). For example, it could be useful if one could ask for the sequence of all genes with a putative signal or anchor sequences or GPI attachment signals. It was suggested (wished) that an integrated and fully annotated "malaria information resource" database be developed that covers not only genome information, but immunochemistry, localization, monoclonal antibody data, protection studies, variant sequences, etc. A dedicated knowledgeable malariologist would be required to annotate it and keep it current. Importantly, the malaria research user community should be actively consulted throughout the development process to ensure that this resource best meets their needs. In fact, the point was made that people will want to be involved in the process of putting together information to make an effective database. The availability of links to appropriate literature was also suggested, especially to facilitate entry of newcomers into malaria research. The presence of mirror sites in different countries was also deemed important.
The yeast (SacchDB) database at <http://genome-www.stanford.edu> was strongly recommended as an excellent model database for cross-communication. The integrated database of the C.elegans genome project was also mentioned as one worth emulating.
F. How can comparative genomic approaches facilitate our understanding of malaria genome information? How can "genomics" be used to obtain information beyond that which can be obtained from "pre-genomic" approaches?
1. Genomics as a science is relatively new and there is clearly a lot to be learned from the many genome projects being conducted today, which could assist investigators in understanding the information contained in malaria genomes.
2. Information from malaria genome project efforts should be linked with information derived from other microbial genome projects. These links should be made to the public sector and to the private or industrial sector.
3. Available information from both the human host and parasite genomes should be easily comparable to facilitate understanding of the nature of the human host-parasite relationships.
4. Information from malaria vector genome projects should also be integrated as it becomes available.
Comment (summarized and condensed)
It could be very useful if a few people working on other genome projects were invited as guest speakers to the biannual malaria genome meeting. On the other hand, it was noted that "given the parlous state of malaria funding, we will be hard pressed to exploit the possibilities generated by the coding regions alone, without bothering with nebulous concepts of genomics."
Malaria Genome Focus group
Enriqueta C. Bond, Martha G. Peck, Jennifer Russo Wortman, Stephanie L. James, Michael Gottlieb, Carole A. Long, Dyann F. Wirth and Mary R. Galinski.
Participants at the April 7th Meeting:
Ripley Ballou, John W. Barnwell, John B. Dame, Mary R. Galinski, Michael Gottlieb, Mark S. Guyer, Stephen L. Hoffman, Stephanie L. James, Altaf A. Lal, Michael Lanzer, Carole A. Long, Wilbur K. Milhous, Robert H. Nagel, Kathryn Nason-Burchenal, Pradip Rathod, Jennifer Russo Wortman, Akhil B. Vaidya, Sara Volkman (Rapporteur), Richard Warren, Andrew P. Waters, Tom E. Wellems, and Dyann F. Wirth (Chair).
John Adams, Ross Coppel, Alan Cowman, Alister Craig, Carter Diggs, Richard Hymen, Dave Lanar, Chris Newbold, Shobhona Sharma, Carol Sibley, David Walliker, Dieter Weichenhan
Special thanks to many others who also wrote or called to express their general interest in this document.