[Federal Register Volume 64, Number 104 (Tuesday, June 1, 1999)]
[Notices]
[Pages 29321-29325]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 99-13740]


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DEPARTMENT OF HEALTH AND HUMAN SERVICES

Centers for Disease Control and Prevention


National Health and Nutrition Examination Survey III (NHANES) DNA 
Specimens: Guidelines for Proposals To Use Anonymized Samples and 
Proposed Cost Schedule

ACTION: Notice and request for comments.

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SUMMARY: The National Health and Nutrition Examination Survey (NHANES) 
is a program of periodic surveys conducted by the National Center for 
Health Statistics (NCHS) of the Centers for Disease Control and 
Prevention (CDC). Examination surveys conducted since 1960 by NCHS have 
provided national estimates of health and nutritional status of the 
United States civilian non-institutionalized population. To add to the 
large amount of information collected for the purpose of describing the 
health of the population in the most recent survey, white cells were 
collected in NHANES III in anticipation of advances in genetic 
research.
    The cells have been stored and maintained at the Division of 
Environmental Health Laboratory Sciences (DEHLS) at the National Center 
for Environmental Health (NCEH), CDC. The collection of white cells was 
begun because of the significant advances in the rapidly evolving field 
of molecular biology that were occurring during the planning phase of 
this survey.
    Technical advances now make it possible to use these samples for 
genetic analysis. NCHS and NCEH, CDC are making anonymized DNA samples 
from these specimens available to the research community for such 
analyses. No cell lines will be made available.
    The purpose of this notice is to request comments on this program 
and cost schedule. After consideration of comments submitted, CDC plans 
to

[[Page 29322]]

finalize the cost schedule and solicit letters of intent and proposals 
for use of the NHANES III anonymized DNA samples. Please contact Ms. 
Burwell or go to www.cdc.gov/nchswww/about/major/nhanes/nhanes.htm for 
final proposal guidelines and request for letters of intent.
    All interested researchers are encouraged to submit letters of 
intent. No funding is provided as part of this solicitation. DNA 
samples will not be provided to those projects requiring funding until 
the project has received funds. Approved projects that do not obtain 
funding will be canceled. A more complete description of this program 
follows.

DATES:
     Comment Receipt Date: June 30, 1999
     Invitation to submit Letters of Intent: July 14, 1999
     Letter of Intent Receipt Date: September 1, 1999
     Invitation to Submit Proposals: September 22, 1999
     Application Receipt Date: November 3, 1999.
     Scientific Review Date: December 1999.
     Institutional Review Date: January 1999.
     Notification of approval: January 1999.
     Anticipated distribution of samples: February-March 2000.

TO SEND COMMENTS AND FOR INFORMATION: Audrey L. Burwell, MS, Health 
Research Administrator, National Center for Health Statistics, Centers 
for Disease Control and Prevention, 6525 Belcrest Road, Room 1100, 
Hyattsville, MD 20782, Phone: 301-436-7062, 127, FAX: 301-436-4233, E-
Mail: [email protected], Internet: www.cdc.gov/nchswww/about/major/nhanes/
nhanes.htm.

SUPPLEMENTARY INFORMATION: The goals of NHANES are: To estimate the 
number and percent of persons in the U.S. population and designated 
subgroups with selected diseases and risk factors; to monitor trends in 
the prevalence, awareness, treatment and control of selected diseases; 
to monitor trends in risk behaviors and environmental exposures; to 
analyze risk factors for selected diseases; to study the relationship 
between diet, nutrition and health; to explore emerging public health 
issues and new technologies; and, to establish and maintain a national 
probability sample of baseline information on health and nutrition 
status.
    The third National Health and Nutrition Examination Survey (NHANES 
III) began in the Fall of 1988 and ended in the Fall of 1994. The 
survey data were collected, and can be analyzed, in two phases: Phase I 
was conducted from October 1988 to October 1991, and Phase II was 
conducted from October 1991 to October 1994. Both phases are nationally 
representative samples. For details of the sampling design see the Plan 
and Operation of NHANES III (1). This information can be obtained by 
contacting the Data Dissemination Branch, NCHS at 301-436-8500 or from 
the Internet at www.cdc.gov/nchswww/products/catalogs/subjects/nhanes3/
nhanes3.htm.
    Blood specimens were collected from participants as a part of 
NHANES III. Lymphocytes were isolated from the blood collected from 
participants aged 12 years and older and stored frozen in liquid 
nitrogen or as cell cultures immortalized with Epstein-Barr virus and 
frozen at the Molecular Biology Branch of DEHLS, NCEH at the CDC, 
Atlanta GA. DNA is available primarily from cell lines of Phase II 
participants.
    Health information collected in the NHANES III is kept in strictest 
confidence. During the informed consent process, survey participants 
are assured that data collected will be used only for stated purposes 
and will not be disclosed or released to others without the consent of 
the individual or the establishment in accordance with section 308(d) 
of the Public Health Service Act (42 U.S.C. 242m). Although the consent 
form was signed by participants in the survey and participants 
consented to storing specimens of their blood for future research, 
specific mention of genetic research was not included. Given the 
scientific importance of this resource, the NHANES Institutional Review 
Board (IRB) approved making anonymized samples of DNA available to the 
research community for genetic research. For this purpose, an 
anonymized sample is defined as a sample for which no one, including 
CDC staff, are able to link the results of the genetic tests back to 
the survey participant (2). NCHS and NCEH will anonymize the samples 
for each request.
    All proposals for testing of anonymized NHANES III DNA samples will 
be evaluated by a Genetics Technical Panel (composed of 8-10 
scientists; 40 percent from CDC; 30 percent from other Federal agencies 
and 30 percent non-government scientists) for scientific merit and by 
the NHANES IRB to assure that anonymity will be maintained, as well as 
for other human subjects concerns. The NHANES IRB review will be 
conducted, even though the investigator may have received review by 
their home institution. The NHANES IRB must review all projects because 
determination of anonymity required by this proposal review process can 
only be accomplished by the NHANES staff who have access to 
confidential records. Projects recommended for approval by the Genetics 
Technical Panel and the IRB will be submitted to the Director of NCHS 
for verification that all appropriate reviews have been conducted.
    The Genetics Technical Panel will evaluate the public health 
significance and scientific merit of the proposed research. Scientific 
merit will be judged as to the scientific, technical or medical 
significance of the research, the appropriateness and adequacy of the 
experimental approach, and the methodology proposed to reach the 
research goals. See Criteria for Technical Evaluation of Proposals 
below. The proposer should outline how the results from the DNA 
analysis will be used. Because NHANES is a complex, multistage 
probability sample of the national population, the appropriateness of 
the NHANES sample to address the goals of the proposal will be an 
important aspect of scientific merit. The Genetics Technical Panel will 
assure that the proposed project does not go beyond either the general 
purpose for collecting the samples in the survey, i.e., to determine 
allele frequencies in subgroups of the population, or of the specific 
stated goals of the proposal. The Panel will also review an evaluation 
by the NCHS staff as to whether anonymity can be assured for the 
proposed project.
    Investigators are encouraged to obtain the NHANES III Reference 
Manuals and Reports and NHANES III Public Use Data (on CD-ROM) These 
can be obtained by contacting the Data Dissemination Branch, NCHS at 
301-436-8500 or from the Internet at www.cdc.gov/nchswww/products/
catalogs/subject/nhanes3/nhanes3.htm.
    Because of the complex nature of this Survey, sampling weights are 
used to make national estimates of frequencies. The use of weights, 
sampling frame and methods of assessment of variables included in the 
data tape are likely to affect the proposed research. The Genetics 
Technical Panel will review the analysis plan and evaluate whether the 
proposal is an appropriate use of the NHANES III population.
    Due to the design of NHANES III, the DNA samples are not suitable 
for family studies. On average, NHANES III sampled 2 individuals from 
each household in Phase II. The relationship

[[Page 29323]]

between individuals is not available on the data file. If the 
investigator requires strict assurance that only one subject per 
household is included among the samples, the investigator should 
request only one subject per household, and estimate statistical cell 
sizes by dividing the results from cross tabulations by 2 (see Special 
Studies procedures below). In this instance, the NCHS staff can use the 
confidential household code (not available on public use tapes) to 
select one subject per household for approved projects.
    The provided samples will consist of DNA suitable for use in the 
polymerase chain reaction or other justifiable genetic assessments. No 
cell lines will be made available. Unique, randomized IDs will be 
assigned to each set of DNA samples.
    Two types of proposals will be evaluated: (a) Those that aim to 
describe allele frequencies which require only basic demographic 
information (age, race, and gender) linked to the samples or (b) those 
where additional co-variates from the NHANES data base are required 
(special studies).

Age-Race-Gender Studies

    To facilitate proposal preparation of allele frequency research, 
NCHS will make available the following data with the DNA sample: age in 
10-year age groups, race-ethnicity (white, black, Mexican-American), 
gender, mean sample weights for each demographic group and the average 
design effect. These proposals, therefore, do not need to provide an 
analysis of NHANES III data to support the anonymization scheme 
proposed. These data have sufficient sample sizes in each category (the 
smallest age, race/ethnicity, gender statistical cell contains 62 
persons) to preserve anonymity. To further preserve anonymity, only 80 
percent of the subjects in each statistical cell will be used.
    Proposals submitted for this review will be limited to those 
requesting samples from within these domains for the identification of 
the frequency of the genes in the population. These proposals must 
address all criteria except for the verification that anonymization can 
be achieved. Because of the limited data associated with the genetic 
result, a shortened (2-3 pages) proposal is acceptable as long as each 
of the criteria below are addressed.

Special Studies (Requests for Additional Variables)

    Include a list of demographic and clinical variables and specify 
recoding schemes, if appropriate, that the principal investigator would 
like to have linked to the DNA samples to meet the objectives of the 
study. The combined information on all variables provided to the 
investigator by CDC must not constitute a unique set of values that 
could link the DNA samples with participant data on the NHANES III 
public use data set. Investigators should obtain the NHANES III Public 
Use Data and should verify prior to submission that anonymity is 
achievable with the requested set of variables. To obtain the NHANES 
III Public Use Data contact the Data Dissemination Branch, NCHS at 301-
436-8500 or from the Internet at www.cdc.gov/nchswww/products/catalogs/
subject/nhanes3/nhanes3.htm.
    Recoding is required for continuous variables and may be required 
for integral variables to assure anonymity. A cross tabulation of all 
requested variables must be provided, with demonstration that there are 
at least five individuals in each statistical cell of that cross 
tabulation. Because the DNA specimens are primarily available on phase 
II subjects, these analyses should be run using phase II subjects only 
(SDPPHASE=2). (Household codes are confidential data. Therefore, if 
only 1 individual per household is to be included in the protocol, the 
investigator can estimate the sample size per statistical cell by 
halving the cross tabulation results. For instance, if only 1 
individual per household is requested, the minimum statistical cell 
size of the cross tabulation should be 10 subjects.) From each 
statistical cell, either 2 subjects or 20 percent of the subjects of 
the cell, whichever is larger, will be deleted from the pool of samples 
sent to the investigator. The DNA samples which are sent to the 
investigator will be selected by the NCHS staff at random from the 
domain.
    All protocols, either for the age-race-gender studies or the 
special studies, will be reviewed by NCHS staff for anonymity and must 
be approved by the Genetics Technical Panel and the NHANES IRB. The 
anonymity of all sample requests, when linked with demographic and 
clinical data, will be verified prior to release to the investigator.
    Applicants may request a project period of up to 3 years. At the 
end of the project period, any unused samples must be returned to the 
Bank in accordance with instructions that will be provided. Extensions 
to the period of performance may be requested.

Procedures for Letter of Intent

    After consideration of comments on the program and the cost 
schedule, NCHS plans to make requests for letters of intent. This 
request will be announced on the NHANES web site by July 14, 1999: 
Internet: www.cdc.gov/nchswww/about/major/nhanes/nhanes.htm. The letter 
of intent is required to enable CDC to plan the review more 
efficiently, evaluate the number and magnitude of the requests, and to 
assess the capacity of the DNA Bank to fulfill requests.
    Investigators from both the extramural and the CDC intramural 
research communities must submit letters of intent. The letter should 
be no more than two pages and include: a descriptive title of the 
overall proposed research, the name, address and telephone number of 
the Principal Investigator, a list of key investigators and their 
institution(s), one paragraph on the background for the proposal, the 
genetic assessments proposed, a list of proposed variables, and an 
estimate of the number of samples that would be requested. The 
background should state concisely the importance of the research in 
terms of the broad, long-term objectives and public health relevance.
    If the total number of proposals and samples requested from all the 
letters of intent exceed the handling capacity of the DNA Specimen 
Bank, a determination of priorities will be made by the Genetics 
Technical Panel using the letters of intent. Priority will be based on 
the public health importance of the proposed research and the 
prevalence of the health outcome(s) targeted by the research.
    All investigators will be notified as to whether they should submit 
a full proposal based on review of the letters of intent.
    Letters of intent should be submitted by September 1, 1999, to: 
Audrey L. Burwell, MS, Health Research Administrator, National Center 
for Health, Statistics, Centers for Disease Control and Prevention 6525 
Belcrest Road, Room 1100, Hyattsville, MD 20782, Phone: 301-436-7062, 
127, FAX: 301-436-4233, E-Mail: [email protected].

Procedures for Proposals

    After notification by NCHS that a proposal for use of samples and 
data from the NHANES III DNA-Bank can be submitted, the investigator 
should use the following format: Proposals are limited to a maximum of 
5 single-spaced typed pages, excluding figures and tables, using 10 cpi 
type density. If a proposal is approved, the title, specific aims, 
name, and phone number of the author will be maintained by NCHS and 
released if requested. Unapproved proposals will be returned to the 
investigator and will not be maintained

[[Page 29324]]

by NCHS. The cover of the proposal should include the name, address, 
and phone number and E-mail address, if available, of the principal 
investigator (PI) and the name of the institution where the DNA 
analysis will be done. The cover page should be signed by the 
responsible institution representative.
    The Criteria for Technical Evaluation of Proposals section at the 
end of this announcement and the following information should be used 
to develop the proposal content.
    1. Specific Aims--List the broad objectives; describe concisely and 
realistically what the research is intended to accomplish, and state 
the specific hypotheses to be tested.
    2. Background and Public Health Significance--Briefly describe in 
1-2 pages the background of the proposal, identifying gaps in knowledge 
that the project is intended to fill. State concisely the importance of 
the research in terms of the broad, long-term objectives and public 
health relevance including a discussion of how the results will affect 
health policy or further scientific knowledge. The proposal should 
include a discussion of the potential clinical significance of the 
results and whether there is definitive evidence that results of the 
genetic test would provide grounds for medical intervention.
    3. Research Design and Methods--Describe the research design and 
the procedures to be used. A detailed description of laboratory methods 
must be included with references. If non-standard methods will be used, 
discuss how the method is more appropriate than current methods or that 
there are no standard methods to accomplish the task. Laboratory 
quality control should be described. Include a justification for 
determination of sample size or a power calculation. A detailed 
description and justification of any sample design whether a random 
subsample or case control design, must be given. The program will 
evaluate the endpoints assessed in these projects to determine whether 
the projects are consistent with the mission of the NHANES program. 
Further, the program and IRB are concerned with the possible breach of 
anonymity due to the determination of a large number of genetic 
analyses in individual research studies. The specific concern is that, 
when large numbers of genetic findings are available that are 
potentially correlated with other NHANES III data, the investigator or 
program staff might be able to inadvertently identify specific 
subjects. Therefore, if several genetic analyses are proposed, the 
investigator must discuss the potential for linking the findings with 
NHANES III data on public use tapes that was not requested as part of 
the proposal. A list of variables requested should be included. A 
crosstabulation demonstrating the recoding and resulting statistical 
cell sizes should be included for special studies.
    4. Qualification of Investigators--A brief description of the 
Principal Investigator's expertise in the proposed area should be 
provided, including publications in this area within the last three 
years. A representative sample of earlier publications may be listed as 
long as this section does not exceed two pages.
    5. Funding--The source and status of the funding to perform the 
requested laboratory analysis should be included. Investigators will be 
responsible for the cost of processing and shipping the samples. At 
this time the cost per DNA specimen is $38.00. The basis for the cost 
structure is in the last section of this document. Reimbursement for 
the samples will be collected before the samples are released.

Requirements for the Inclusion of Women and Racial and Ethnic 
Minorities in Research

    In NHANES III, race/ethnicity was defined by self-report as non-
Hispanic white, non-Hispanic black, or Mexican American. Individuals 
who did not self-select into these categories were classified as 
``other'' . If the proposal excludes one or more race/ethnic groups or 
a gender, this exclusion must be justified.
    The CDC is also sensitive to the stigmatization of racial/ethnic 
specific populations through inappropriate reporting and interpretation 
of findings. For all proposals that request information on race/
ethnicity for the samples selected, the investigator should indicate 
the reason for analyzing race/ethnicity and how the results will be 
interpreted.

Submission of Proposals

    Investigators who are invited to submit proposals should send the 
original written proposal and 20 copies to: Audrey L. Burwell, MS, 
Health Research Administrator, National Center for Health Statistics, 
6525 Belcrest Rd., Rm 1100, Hyattsville, MD 20782, Phone: (301) 436-
7062, 127, FAX: (301) 436-4233, E-Mail: [email protected], Attention: NHANES 
III Genetic Testing Program.

Criteria for Technical Evaluation of Proposals

    The following criteria will be used for technical evaluation of 
proposals.
    1. Background and Public Health Significance: The public health 
significance, scientific merit and practical utility of the assay. The 
proposer has conveyed how the results will be used and the relationship 
of the results to the data already collected in NHANES III. Proposer 
addresses how they will use results of the DNA analyses. The analyses 
are consistent with the NHANES mission and the health status variables 
will be evaluated appropriately given the methods of data collection in 
NHANES III.
    2. Research Design and Methods: The sampling scheme must be 
described and address its relationship to the NHANES III design. Power 
calculations for a subsample must be included. A list of variables 
requested with the recoding schemes are included. A cross-tabulation is 
provided if the investigator requests a special set of variables. A 
detailed description of the laboratory methods is included. If a non-
standard laboratory method is to be used, its advantage over existing 
methodology is adequately discussed. The characteristics of the 
laboratory assay, such as reliability, validity, and ``state-of-the-
art'', must be included with appropriate references. The potential 
difficulties and limitations of the proposed procedures are discussed. 
The volume of DNA and the number of samples required are specified. 
Adequate methods for handling and storage of samples must be addressed. 
The laboratory has demonstrated the capability for handling the 
workload requested in the proposal.
    3. Discussion regarding the race/ethnicity and gender variables: If 
either race/ethnicity or gender are used to restrict the sample, the 
proposal gives a clear and compelling rationale for this restriction. 
On the other hand, if the race/ethnicity variable is requested, the 
proposer indicates the reason for analyzing race/ethnicity and how the 
results will be interpreted.
    4. Qualifications: A brief description of the requestor's expertise 
in the proposed area is provided including publications in this area 
within the last three years. A representative sample of earlier 
publications may be listed as long as this section does not exceed two 
pages.
    5. Anonymity: NCHS determination of anonymity has been reviewed and 
found to be adequate.
    6. Period of performance--The project period should be specified. 
The period may be up to three years. At the end of the project period, 
any unused samples must be returned to the NHANES DNA Specimen Bank in 
accordance with instructions from the DEHLS.

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Extensions to the period of performance may be requested.

Approved Proposals

    NCHS/NCEH will provide a data file with the requested recoded 
variables and a randomly assigned unique identification number that is 
linked to the DNA specimen. No record connecting the new number with 
the original identification number will be kept after the samples have 
been sent. These samples can not be traced to any files maintained by 
NCHS.

Agency Agreement

    A formal signed agreement in the form of a Materials Transfer 
Agreement (MTA) with individuals who have projects approved will be 
completed before the release of the samples. This agreement will 
contain the conditions for use of the DNA as stated in this document 
and as agreed upon by the investigators and CDC. A key component of 
this agreement is that no attempt will be made to link the results of 
the proposed research to any other data, including, but not limited to, 
the NHANES III public use data set. Also, the investigator agrees that 
the samples can not be used for commercial purposes.

Progress Reports

    A progress report will be submitted annually. NHANES IRB 
continuation reports are also required annually.

Disposition of Results and Samples

    No DNA samples provided can be used for any purpose other than 
those specifically requested in the proposal and approved by the 
Genetic Technical Panel and the NHANES IRB. No sample can be shared 
with others, including other investigators, unless specified in the 
proposal and so approved. Any unused samples must be returned to the 
Bank upon completion of the approved project. Researchers requesting 
DNA samples for age-race-gender studies and special studies will be 
required to provide NCHS with the results of all DNA tests performed 
for each anonymized sample. These results, once returned to NCHS, will 
be part of the public domain. Therefore, ample time will be given to 
the investigator to publish results prior to reporting the results to 
NCHS.

Proposed Cost Schedule For Providing NHANES III DNA Specimen Bank

    A nominal processing fee of $38.00 is proposed for each sample 
received from the NHANES III DNA Specimen Bank. The costs are 
determined both for NCEH and NCHS and include the physical materials 
needed to process the samples at the NCEH laboratory as well as the 
materials to process the requests for samples at NCHS. These costs are 
inclusive of the staff needed for these activities at each Center. The 
fee is estimated to cover the costs of processing, handling and 
preparing the samples in accordance with the detailed requirements of 
the investigators. These costs were based on an assumption that NCEH 
and NCHS will receive and process 15 proposals in a year each 
requesting 1000 samples as shown in the table below.
    The materials listed are for the recurring laboratory costs to 
dispense and prepare the samples for shipping; the computer software 
needed for the Web page and advertisements in scientific journals. 
Labor costs are based on the need for microbiologists, a proposal 
administrator and computer programmers for NCHS and NCEH to maintain 
the data bases and verify anonymity. Technical panel travel and 
expenses are based on the panel meeting twice a year. The space 
estimate is based on acquiring storage and aliquoting space.

------------------------------------------------------------------------
 
------------------------------------------------------------------------
        Total Costs Per Sample if 15 Requests for 1,000 Samples
------------------------------------------------------------------------
Materials..................................................        $1.90
Labor......................................................        22.00
Panel Travel/Expenses......................................         2.69
Space......................................................         0.97
Subtotal...................................................        27.56
NCHS overhead (15%)........................................         4.12
Subtotal...................................................        31.68
CDC/FMO overhead (20%).....................................         6.32
                                                            ------------
    Total..................................................     * 38.00
------------------------------------------------------------------------
* Shipping costs are not included in the $38.00 processing fee. These
  costs must also be paid by the investigator.

    Comments are solicited on the proposed cost schedule. Comments are 
due by June 30, 1999.

Send Comments and for Information

Audrey L. Burwell, MS, Health Research Administrator, National Center 
for Health Statistics, Centers for Disease Control and Prevention, 6525 
Belcrest Road, Room 1100, Hyattsville, MD 20782, Phone: 301-436-7062, 
127, FAX: 301-436-4233, E-Mail: [email protected]

References

    1. Plan and Operation of the Third National Health and Nutrition 
Examination Survey, 1988-94. National Center for Health Statistics. 
Vital Health Stat (32) 1994.
    2. Clayton EW, Steinberg KK, Khoury MJ, et al. Informed consent 
for genetic research on stored tissue samples. JAMA 1995;274:1786-
1792.

    Dated: May 25, 1999.
Joseph R. Carter,
Acting Associate Director for Management And Operation, Centers for 
Disease Control and Prevention.
[FR Doc. 99-13740 Filed 5-28-99; 8:45 am]
BILLING CODE 4163-18-P