DEVELOPMENT OF HIGH-YIELD TECHNOLOGIES FOR ISOLATING EXFOLIATED CELLS IN BODY 
FLUIDS
 
Release Date:  November 15, 2000  

RFA: CA-01-016

National Cancer Institute

Letter of Intent Receipt Dates:  March 6, 2001    
Application Receipt Dates: April 10, 2001

PURPOSE

The purpose of this Request for Application (RFA) is to develop novel 
technologies for capturing, enriching, and preserving exfoliated abnormal 
cells in body fluids or effusions and to develop methods for concentrating the 
enriched cells for biomarker studies. In body fluids, such as sputum, the 
number of exfoliated tumor cells is often small compared to the number of non-
neoplastic cells. Therefore, the detection of exfoliated abnormal cells by 
routine cytopathology is often limited because few atypical cells may be 
present in the specimen. Furthermore, there may be difficulty in separating 
dysplastic cells from non-specific reactive changes and degenerating cells or 
variation in diagnostic criteria. Furthermore, exfoliated cells are frequently 
contaminated with normal cells, bacteria, and other cellular debris, which 
makes molecular analysis difficult without physical separation of the 
neoplastic cells. Thus, the development of enrichment methods is a 
prerequisite for the routine detection of small numbers of exfoliated cells 
and small amounts of subcellular materials in biological fluids for molecular 
analysis.

This Request for Application (RFA) must be read in conjunction with the 
OMNIBUS SOLICITATION OF THE NATIONAL INSTITUTES OF HEALTH, SMALL BUSINESS 
INNOVATION RESEARCH (SBIR) and SMALL BUSINESS TECHNOLOGY TRANSFER (STTR) GRANT 
APPLICATIONS (PHS 2001-2).  PHS-2001-2 will be issued in January 2001.  
Therefore, applicants may want to refer to PHS 2000-2 
(http://grants.nih.gov/grants/funding/sbir.htm) as a general reference until 
the Calendar Year 2001 SBIR/STTR Omnibus Solicitation is released
 
All of the instructions within the Omnibus Solicitation apply with the 
following exceptions:
  
o  Special receipt dates    
o  Initial review convened by the NCI Division of Extramural Activities    
o  Additional review considerations

This RFA will utilize the Small Business Innovation Research (SBIR) and Small 
Business Technology Transfer (STTR) mechanisms, but will be run in parallel 
with a Program Announcement of identical scientific scope PAR-01-019 that will 
utilize the exploratory/developmental (R21) grant mechanism. PAR-01-019 is 
available at (see http://grants.nih.gov/grants/guide/pa-files/PAR-01-
019.html).

RESEARCH OBJECTIVES 

Background

The most common human tumors arise from epithelial surfaces (e.g. colon, lung, 
prostate, oral cavity, esophagus, stomach, uterine cervix, bladder). Their 
development often becomes apparent when tumor cells exfoliate spontaneously 
into sputum, urine, or even into various effusions.  The molecular and genetic 
abnormalities within these exfoliated cells could be used to detect and 
identify precancerous lesions or very early stage cancer if highly sensitive 
technologies were clinically available to identify the few abnormal cells 
among millions of normal cells.   For example, detection of widespread 
microsatellite instability (MSI), as demonstrated by expansion or deletion of 
repeat elements of DNA, may be adapted for exfoliated cells in general.  With 
the advent of PCR-based detection of DNA from rare neoplastic cells in body 
fluids, mutations have been detected in ras genes from the stool of patients 
with colorectal cancer, and in p53 from the urine of patients with bladder 
cancer and in the sputum of patients with lung cancer.  As these assays are 
complex and technically challenging, they depend on the development of novel 
technologies for isolating and enriching exfoliated cells.  

Abnormal exfoliated cells can be routinely identified by cytologic examination 
of brushings and fluids, for instance, from bronchi, pancreatic ducts, voided 
urine and tapping of effusions. Currently fluids are usually processed by 
centrifugation or membrane filtration. However, the detection of abnormal 
exfoliated cells, for instance, cancer cells by routine cytopathological 
examination may be limited because the number of abnormal cells may be very 
small compared to the number of normal cells. Alternatively, the cellular and 
nuclear changes in abnormal cells may be minimal compared to normal cells. 
This is particularly true of urine cytology, where many low-grade papillary 
lesions are often missed on cytological examination.  New PCR-based 
technologies may substantially enhance the  sensitivity, but current 
technologies for isolating exfoliated cells are too cumbersome to be of 
practical utility. 
Finding molecular and genetic biomarkers of early cancer represents an 
extraordinary opportunity for the National Cancer Institute (NCI) and is 
particularly important in detecting the emergence of precancerous cell 
populations.  In these earliest stages of neoplastic development, lesions 
should be amenable to complete eradication.  This has been well demonstrated 
in cervical neoplasia, where screening for brushed exfoliated cells has 
resulted in a 70% or greater reduction in cervical cancer mortality. Brushing 
of the esophagus or stomach is commonly performed during endoscopic 
examination.

Goals and Scope

In pursuit of these goals, the NCI invites applications, which address the 
following areas: 
o  development of novel technologies for identifying abnormal exfoliated cells 
in body fluids; 
o  development of novel technologies for capturing, enriching, and preserving 
abnormal exfoliated cells in body fluids; 
o  development of enrichment methods for the isolation of tumor cells;
o  development of sensitive, high-throughput molecular, cytomorphometric, 
immunologic, and other relevant technologies to isolate tumor cells in 
malignant effusions for detection of low tumor burden and to help distinguish 
reactive cells from tumor cells. 

Applicants should not only address the technology of enriching and isolating 
exfoliated cells, but they should also address their viability and usefulness 
for cytologic and molecular studies.

MECHANISM OF SUPPORT 

Support for the RFA is through the SBIR and STTR mechanisms, which are set-
aside programs. Awards will be administered under NIH grants policy stated in 
the NIH Grants Policy Statement, NIH publication 99-8 October 1998.

Applications can be submitted for support as Phase I STTR (R41) or Phase I 
SBIR (R43) grants: Phase II STTR (R42) or Phase II SBIR (R44) grants; or under 
the SBIR/STTR  FAST-TRACK option as described in the OMNIBUS SOLICITATION.  
Phase II applications in response to this RFA will only be accepted as 
competing continuations of previously funded NIH Phase I SBIR/STTR awards.  
The Phase II proposal must be a logical extension of the Phase I research.

Information on the FAST-TRACK process and the OMNIBUS SOLICITATION are 
available at: http://grants.nih.gov/grants/funding/sbirsttr1/index.htm

FUNDS AVAILABLE

$ 1 M to support up to 6 Phase I and/or Phase II applications under the 
SBIR/STTR set-aside  funding mechanism are available. 

Although the financial plans of the NCI provide support for this program, 
awards pursuant to this RFA are contingent upon the availability of funds and 
the receipt of a sufficient number of meritorious applications.   At this 
time, it is not known if competing renewal applications will be accepted 
and/or if this RFA will be reissued.

ELIGIBILITY REQUIREMENTS

Eligibility requirements are described in the OMNIBUS SOLICITATION.  Any small 
business, independently owned by United States citizens and located in the 
United States, may apply.

INQUIRIES

Direct inquiries regarding programmatic issues to:

Sudhir Srivastava, Ph.D., M.P.H.
Division of Cancer Prevention
National Cancer Institute
EPN, Room 330F
6130 Executive Boulevard
Rockville, MD 20852
Telephone:  (301) 435-1594
FAX:  (301) 402 -0816
Email:  ss1a@nih.gov
 
Direct inquiries regarding review matters to:

Ms. Toby Friedberg
Division of Extramural Activities
National Cancer Institute
6116 Executive Boulevard, Room 8109, MSC 8329
Bethesda, MD  20892-8329
Rockville, MD  20852 (for express/courier service)
Telephone: (301) 496 -3428
FAX: (301) 402-0275 
Email: tf12w@nih.gov

Direct inquiries regarding fiscal matters to: 
 
Ms. Kathleen Shino 
Grants Administration Branch
National Cancer Institute 
6120 Executive Boulevard, Room 243, MSC 7150 
Bethesda, MD  20892-7150 
Rockville, MD 20852 (express courier)
Telephone:  (301) 496-8635 
Fax:  (301) 496-8601
Email:  shinok@gab.nci.nih.gov

LETTER OF INTENT

Prospective applicants are asked to submit, by the date listed on the first 
page of this RFA, a letter of intent that includes a descriptive title of the 
proposed research, the name, address, and telephone number of the Principal 
Investigator, the identities of other key personnel and participating 
institutions, and the number and title of the RFA in response to which the 
application may be submitted.  Although a letter of intent is not required, is 
not binding, and does not enter into the review of a subsequent application, 
the information that it contains allows IC staff to estimate the potential 
review workload and plan the review.
 
The letter of intent is to be sent to Dr. Sudhir Srivastava by the letter of 
intent receipt date listed.

SCHEDULE
 
Letter of Intent Receipt:          March 6, 2001
Application Receipt Date:          April 10, 2001
Peer Review Date:                  June 2001
Review by NCAB Advisory Board:     September 2001
Earliest Anticipated Start Date:   December 2001

APPLICATION PROCEDURES

OMNIBUS SOLICITATION for both the SBIR and STTR programs are available 
electronically through the NIH, Office of Extramural Research small Business 
Funding Opportunities web site at 
http://grants.nih.gov/grants/funding/sbirsttr1/index.htm.  Only a limited 
number of hard copies will be available, and they may be obtained from the PHS 
SBIR/STTR Solicitation Office, phone (301) 206-9385; FAX (301) 206-9722; email 
a2y@cu.nih.gov.  Helpful information for preparation of the application can be 
obtained: http://grants.nih.gov/grants/funding/sbir.htm.  Applications are to 
be submitted on the grant application form PHS 6246-1 (1/99) (SBIR) and PHS 
6246-3 (STTR) (3/99) (in a single document) located in the back pages of the 
OMNIBUS SOLICITATION, and will be accepted at the application deadlines as 
indicated on the first page of this document.  THE TITLE AND NUMBER OF THIS 
RFA MUST BE TYPED IN LINE 2 ON THE FACE PAGE OF THE APPLICATION. 

The OMNIBUS SOLICITATION give the normal levels of support and period of time 
for SBIR and STTR Phase I and II awards.  However, these award levels are 
guidelines and not ceilings.  Therefore, larger budgets with longer periods of 
time may be requested if required to complete the proposed research.  As 
stated under MECHANISM OF SUPPORT section, Phase I applications submitted in 
response to this RFA can have a project period of up to two years and a budget 
not to exceed $100,000 per year direct cost excluding subcontractor indirect 
costs. 

The second year of the Phase I budget should be included on the Budget 
Justification page, using categorical totals if costs deviate significantly 
from the first year of the budget, with narrative justifications for the 
increase(s).  If the second year simply escalates due to cost of living 
factors, a statement to that effect with the escalation factor should be 
included rather than categorical totals. Phase II applications submitted to 
this RFA that exceed the normal levels of support as stated in the OMNIBUS 
SOLICITATION must be strongly justified. The total duration (Phase I and Phase 
II application) cannot exceed four years.

In order to apply for the FAST-TRACK option, applications for both Phase I and 
Phase II must be submitted together according to the instructions for FAST 
TRACK applications as described in the OMNIBUS SOLICITATION.  The Phase I 
application must specify clear, well-defined quantifiable milestones that 
should be achieved prior to Phase II funding. Milestones should be located in 
a separate section at the end of the Research Plan of the Phase I and should 
be indicated in the Table of Contents.  Failure to provide measurable 
milestones and sufficient detail may be sufficient reason for the peer review 
committee to exclude the Phase II application from FAST-TRACK review. If so, 
at a later date, the applicant may apply for Phase II support through normal 
application procedures.  Such applications will be reviewed by a standard 
Study Section of the Center for Scientific Review or by a special review group 
convened in response to a re- issuance of this RFA, if applicable.  

An additional requirement of the FAST-TRACK mechanism is the Product 
Development Plan.  The small business must submit a concise Product 
Development Plan (limited to ten pages) as an Appendix to the Phase II 
application addressing the four areas described in the instructions for FAST-
TRACK applications in the OMNIBUS SOLICITATION.  In the event that an 
applicant feels that technology is too proprietary to disclose, applicants at 
a minimum should  provide a demonstration (e.g., results) of the capabilities 
of the proposed technology.  

Submit a signed, typewritten original of the application, including the 
Checklist, and one signed, photocopy, in one package to:
 
Center for Scientific Review
National Institutes of Health
6701 Rockledge Drive, Room 1040 - MSC 7710
BETHESDA, MD  20892-7710
BETHESDA, MD  20817 (for express/courier service)

To expedite the review process, at the time of submission, send one additional
copy of the application to:  

Ms. Toby Friedberg
Division of Extramural Activities
National Cancer Institute
6116 Executive Boulevard, Room 8109, MSC 8329
Bethesda, MD  20892-8329
Rockville, MD  20852 (for express/courier service)
Telephone:  (301) 496-3428
FAX:  (301) 402-0275

Applications must be received by the receipt date listed at the beginning of 
this RFA.

REVIEW CONSIDERATIONS

Upon receipt, applications will be reviewed for completeness by CSR and 
responsiveness by the National Cancer Institute. Incomplete and/or non-
responsive applications will be returned to the applicant without further 
consideration.  

Applications that are complete and responsive to the RFA will be evaluated for 
scientific and technical merit by an appropriate peer review group convened by 
the Division of Extramural Activities of the NCI in accordance with the review 
criteria stated below.  As part of the initial merit review, all applications 
will receive a written critique and undergo a process in which only those 
applications deemed to have the highest scientific merit generally the top 
half of the applications under review will be discussed, assigned a priority 
score, and receive a second level review by the National Cancer Advisory Board 
(NCAB).
    
Review Criteria.

Review criteria are described in the NIH Omnibus Solicitation and available on 
the web at the following URL address: 
http://grants.nih.gov/grants/funding/sbirsttr1/index.htm 

In addition to the standard review criteria as described in the NIH OMNIBUS 
SOLICITATION, the reviewers will comment on the six following aspects of the 
application in their written critiques in order to judge the likelihood that 
the proposed research will have a substantial impact on the pursuit of these 
goals.  Each of these criteria will be addressed and considered by the 
reviewers in assigning the overall score weighting them as appropriate for 
each application. Note that the application does not need to be strong in all 
categories to be judged likely to have a major scientific impact and thus 
deserve a high priority score.  For example, an investigator may propose to 
carry out important work that by its nature is not innovative but is essential 
to move a technology forward. 

Significance. Does this study address the objectives of this RFA? If the aims 
of  the application are achieved, how will this project enable the 
technological application of exfoliated cells?  What will be the effect of 
these studies on the concepts or methods that drive the non-invasive 
approaches to cancer detection?  To what degree does the technology support 
the needs of the targeted research community?
  
Approach.  Are the conceptual framework, design, methods, and analyses 
adequately developed, well-integrated, and appropriate to the aims of the 
project?  Does the applicant acknowledge potential problem areas and consider 
alternative tactics? What is the time frame for developing the proposed 
technologies and suitability of this time frame for meeting the scientific 
community’s needs?  How easy will it be to use the proposed technology?  Are 
the plans for proposed technology dissemination adequate?  
  
Milestones.  How appropriate are the proposed milestones against which to 
evaluate the demonstration of feasibility for transition to the Phase II 
development phase?
  
Innovation.  Does the project employ novel concepts, approaches or method? Are 
the aims original and innovative? Does the project challenge existing 
paradigms or develop new methodologies or technologies? What is the throughput 
and cost effectiveness of the proposed technology?  What additional uses can 
be projected for the proposed technology?

Investigator.  Is the investigator appropriately trained and well suited to 
carry out this work?  Is the work proposed appropriate to the experience level 
of the principal investigator and other researchers (if any)?
  
Environment.  Does the scientific environment in which the work will be done 
contribute to the probability of success?  Do the proposed experiments take 
advantage of unique features of the scientific environment or employ useful 
collaborative arrangements? Is there evidence of institutional support?
        
The initial review group will also examine: the appropriateness of the 
proposed project budget and duration; the adequacy of plans to include both 
genders and minorities and their subgroups, and children as appropriate for 
the scientific goals of the research and plans for the recruitment and 
retention of subjects; the provisions for the protection of human and animal 
subjects; and the safety of the research environment.
    
AWARD CRITERIA

Applications will compete for available funds with all other recommended SBIR 
and STTR applications.  Funding decisions for Phase I or Phase II applications 
will be based on quality of the proposed project as determined by peer review, 
availability of funds, and program priority.

FAST-TRACK, Phase II applications may be funded following submission of the 
Phase I progress report and other documents necessary for continuation.  Phase 
II applications will be selected for funding based on the initial priority 
score, NCI’s assessment of the Phase I progress and determination that Phase I 
goals were achieved, the project’s potential for commercial success, and the 
availability of funds.

INCLUSION OF WOMEN AND MINORITIES IN RESEARCH INVOLVING HUMAN SUBJECTS

It is the policy of the NIH that women and members of minority groups and 
their subpopulations must be included in all NIH supported biomedical and 
behavioral research projects involving human subjects, unless a clear and 
compelling rationale and justification is provided that inclusion is 
inappropriate with respect to the health of the subjects or the purpose of the 
research.  This policy results from the NIH Revitalization Act of 1993 
(Section 492B of Public Law 103-43).
 
All investigators proposing research involving human subjects should read the 
UPDATED "NIH Guidelines for Inclusion of Women and Minorities as Subjects in 
Clinical Research," published in the NIH Guide for Grants and Contracts on 
August 2, 2000  (http://grants.nih.gov/grants/guide/notice-files/NOT-OD-00-
048.html); a complete copy of the updated Guidelines are available at 
http://grants.nih.gov/grants/funding/women_min/guidelines_update.htm: The 
revisions relate to NIH defined Phase III clinical trials and require: a) all 
applications or proposals and/or protocols to provide a description of plans 
to conduct analyses, as appropriate, to address differences by sex/gender 
and/or racial/ethnic groups, including subgroups if applicable; and b) all 
investigators to report accrual, and to conduct and report analyses, as 
appropriate, by sex/gender and/or racial/ethnic group differences.

INCLUSION OF CHILDREN AS PARTICIPANTS IN RESEARCH INVOLVING HUMAN SUBJECTS

It is the policy of NIH that children (i.e., individuals under the age of 21) 
must be included in all human subjects research, conducted or supported by the 
NIH, unless there are clear and compelling scientific and ethical reasons not 
to include them.  This policy applies to all initial (Type 1) applications 
submitted for receipt dates after October 1, 1998.

All investigators proposing research involving human subjects should read the 
“NIH Policy and Guidelines on the Inclusion of Children as participants in 
Research Involving Human Subjects” that was published in the NIH Guide for 
Grants and Contracts, March 6, 1998, and is available at the following URL 
address: http://grants.nih.gov/grants/guide/notice-files/not98-024.html. 

Investigators also may obtain copies of the policy from the program staff 
listed under INQUIRIES.  Program staff may also provide additional relevant 
information concerning the policy.

URLS IN NIH GRANT APPLICATIONS OR APPENDICES 

All applications and proposals for NIH funding must be self-contained within 
specified page limitations.  Unless otherwise specified in a NIH solicitation, 
Internet addresses (URLs) should not be used to provide information necessary 
to the review because reviewers are under no obligation to view the Internet 
sites.  Reviewers are cautioned that their anonymity may be compromised when 
they directly access an Internet site.

HEALTHY PEOPLE 2010

The Public Health Service (PHS) is committed to achieving the health promotion 
and disease prevention objectives of "Healthy People 2010," a PHS led national 
activity for setting priority areas. This RFA, Development of High-Yield 
Technologies for Isolating Exfoliated Cells in Body Fluids, is related to the 
priority area of Cancer. Potential applicants may obtain a copy of "Healthy 
People 2010" at http://www.health.gov/healthypeople/.

AUTHORITY AND REGULATIONS

This program is described in the Catalog of Federal Domestic Assistance No. 
93.393 Cancer Cause and Prevention Research. Awards are made under 
authorization of Sections 301 and 405 of the Public Health Service Act as 
amended (42 USC 241 and 284) and administered under NIH grants policies and 
Federal Regulations 42 CFR 52 and 45 CFR parts 74 and 92. This program is not 
subject to the intergovernmental review requirements of Executive Order 12372 
or Health Systems Agency review.

The PHS strongly encourages all grant recipients to provide a smoke-free 
workplace and promote the non-use of all tobacco products. In addition, Public 
Law 103-227, the Pro-Children Act of 1994, prohibits smoking in certain 
facilities (or in some cases, any portion of a facility) in which regular or 
routine education, library, day care, health care, or early childhood 
development services are provided to children. This is consistent with the PHS 
mission to protect and advance the physical and mental health of the American 
people.

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