Broccoli Sprout Extracts in Healthy Volunteers: A Pilot Study of Nrf2 Pathway Modulation in Oral Mucosa
Broccoli Sprout Extracts in Healthy Volunteers: A Pilot Study of Nrf2 Pathway Modulation in Oral Mucosa
A pilot study has been designed to determine (primary objective) if three brief interventions
 with three oral BSE regimens will alter mRNA biomarkers of Nrf2 pathway signaling, including
 NQO1, GSTs and AKRs, in the oral mucosa of healthy subjects. Quantitative distribution data
 and preliminary effect size for specific Nrf2 targets, as measured in serial buccal cell
 scrapings, will be determined during the course of 3-day exposures to three BSE regimens.
 These data will inform the design of a randomized, phase II chemoprevention trial in patients
 with HPV-negative HNSCC.

 Ten healthy volunteers will be recruited for this pilot study, Age ≥ 18 years, both male and
 female: 1) The non-cancer population presenting to the University of Pittsburgh Eye and Ear
 Institute or the Hillman Cancer Center. This may include patients with benign disease or
 their friends/family members, or friends/family members of patients with cancer; 2)
 Professionals, staff, or students at the University of Pittsburgh.
Intervention 1, GRR-BSE

 If eligible, participants may undergo baseline visit procedures immediately following the
 screening visit (same day). If necessary for logistical purposes, participants may return for
 the baseline visit and initiation of the first BSE intervention any time during the 14 days
 following eligibility assessment. At the baseline visit (day 1), participants will undergo
 baseline blood and buccal cell collection. Blood collection will be performed by a trained
 phlebotomist. Buccal cell collection will be conducted by a trained research coordinator, the
 principal investigator, or a co-investigator. Participants will receive written and verbal
 instructions and an intervention diary for documentation of the BSE drink, details of the
 intervention timeline, including overnight voids, blood and buccal cell collection, and days
 of GRR-BSE consumption. Participants will receive a list of brassica vegetables and
 condiments to avoid during each study intervention. Participants will also receive a list of
 CYP3A4 inducers and inhibitors that must be avoided during the study intervention. For
 overnight voids, participants will be instructed to begin collecting urine at 5 pm, and to
 collect all urine through the first morning void on the following day. The intervention diary
 will have space for participants to record the date and time of each GRR-BSE consumption, and
 space for subjective comments regarding taste and tolerability. Participants will be provided
 with: 1) a collection bottle for overnight urine collection; 2) three frozen GRR-BSE
 beverages in an insulated cooler bag with a frozen blue-ice block; 3) the intervention diary
 and instructions, as stated above.

 Participants will store the frozen GRR-BSE beverages in their home freezer. On the evening of
 day 1, participants will collect a baseline overnight void which will be returned to the
 study team on day 2. Each evening for 3 evenings (days 2-4; between 5pm and 8pm),
 participants will thaw and consume one GRR-BSE beverage. Beverages may be thawed at room
 temperature for 1-2 hours, or in a lukewarm (70-80°F) water bath for 20-30 minutes.
 Participants will record the time of consumption on the intervention diary. AE assessment and
 buccal cell collections will be conducted on days 3, 4, and 5 (between 8 am and 1 pm) by a
 study investigator or trained research coordinator. On the evening of day 4, participants
 will collect an overnight void which will be returned to the study team on day 5 along with
 the intervention diary. Blood collection will occur again on day 5 (between 8 am and 1 pm).

 Participants will then undergo a minimum 3 day washout. Day 5 of the intervention, which does
 not include a GRR-BSE dose, counts as the first day of the washout period.

 Intervention 2, SFR-BSE

 At the baseline visit for intervention 2 (day 1), participants will undergo baseline blood
 and buccal cell collection. Blood collection will be performed by a trained phlebotomist.
 Buccal cell collection will be conducted by a trained research coordinator, the principal
 investigator, or a co-investigator. Participants will receive written and verbal instructions
 and an intervention diary for documentation of the SFR-BSE drink, details of the intervention
 timeline, including overnight voids, blood and buccal cell collection, and days of SFR-BSE
 consumption. Participants will receive a list of brassica vegetables and condiments to avoid
 during each study intervention. Participants will also receive a list of CYP3A4 inducers and
 inhibitors that must be avoided during the study intervention. For overnight voids,
 participants will be instructed to begin collecting urine at 5 pm, and to collect all urine
 through the first morning void on the following day. The intervention diary will have space
 for participants to record the date and time of each SFR-BSE consumption, and space for
 subjective comments regarding taste and tolerability. Participants will be provided with: 1)
 a collection bottle for overnight urine collection, including instructions for collection; 2)
 three frozen SFR-BSE beverages in an insulated cooler bag with a frozen blue-ice block; 3)
 the intervention diary and instructions, as stated above.

 Participants will store the frozen SFR-BSE beverages in their home freezer. On the evening of
 day 1, participants will collect a baseline overnight void which will be returned to the
 study team on day 2. Each evening for 3 evenings (days 2-4; between 5pm and 8pm),
 participants will thaw and consume one SFR-BSE beverage. Beverages may be thawed at room
 temperature for 1-2 hours, or in a lukewarm (70-80°F) water bath for 20-30 minutes.
 Participants will record the time of consumption on the intervention diary. AE assessment and
 buccal cell collections will be conducted on days 3, 4, and 5 (between 8 am and 1 pm) by a
 study investigator or a trained research coordinator. On the evening of day 4, participants
 will collect an overnight void which will be returned to the study team on day 5 along with
 the intervention diary. Blood collection will occur again on day 5 (between 8 am and 1 pm).

 Participants will then undergo a minimum 3 day washout. Day 5 of the intervention, which does
 not include a SFR-BSE dose, counts as the first day of the washout period.

 Intervention 3, SFR-BSE(T)

 At the baseline visit for intervention 3 (day 1), participants will undergo baseline blood
 and buccal cell collection. Blood collection will be performed by a trained phlebotomist.
 Buccal cell collection will be conducted by a trained research coordinator, the principal
 investigator, or a co-investigator. Participants will receive written and verbal instructions
 and an intervention diary for documentation of the SFR-BSE (T) application, details of the
 intervention timeline, including overnight voids, blood and buccal cell collection, and days
 of SFR-BSE (T) application. Participants will receive a list of brassica vegetables and
 condiments to avoid during each study intervention. Participants will also receive a list of
 CYP3A4 inducers and inhibitors that must be avoided during the study intervention. For
 overnight voids, participants will be instructed to begin collecting urine at 5 pm, and to
 collect all urine through the first morning void on the following day. Participants will be
 explicitly instructed that intervention 3 is topical application only. Participants will be
 instructed to take one mouthful (approximately 30 mL or 1/3 of the dose) of the SFR-BSE(T)
 beverage, swish, gargle, and hold in the mouth for 2-3 minutes then spit out. This will be
 repeated twice (for a total of 3 swish, gargle, hold and spit applications) until the
 beverage is gone, aiming for a total exposure time of 6-8 minutes. The intervention diary
 will have space for participants to record the date, time and duration of each SFR-BSE(T)
 topical application, and space for subjective comments regarding taste and tolerability.
 Participants will be provided with: 1) a collection bottle for overnight urine collection,
 including instructions for collection; 2) three frozen SFR-BSE (T) beverages in an insulated
 cooler bag with a frozen blue-ice block; 3) the intervention diary and instructions, as
 stated above.

 Participants will store the frozen SFR-BSE(T) beverages in their home freezer. On the evening
 of day 1, participants will collect a baseline overnight void which will be returned to the
 study team on day 2. Each evening for 3 evenings (days 2-4; between 5pm and 8pm),
 participants will thaw and consume one SFR-BSE beverage. Beverages may be thawed at room
 temperature for 1-2 hours, or in a lukewarm (70-80°F) water bath for 20-30 minutes.
 Participants will record the time and duration of each topical application on the
 intervention diary. AE assessment and buccal cell collections will be conducted on days 3, 4,
 and 5 (between 8 am and 1 pm) by a study investigator. On the evening of day 4, participants
 will collect an overnight void which will be returned to the study team on day 5 along with
 the intervention diary. Blood collection will occur again on day 5 (between 8 am and 1 pm).
Inclusion Criteria:

 1. Age ≥ 18 years. Children will not be recruited into this study because children do not
 develop head and neck cancer.

 2. Both males and females are eligible.

 3. Members of all racial and ethnic groups are eligible.

 4. Smoking and non-smoking people are eligible. The tobacco use assessment form must be
 completed following consent and registration (Appendix B).

 5. No current or former diagnosis of cancer, with the exception of: excised and cured
 non-melanoma skin cancer; or carcinoma in situ of the cervix

 6. No use of chronic prescribed medications which are potent inducers or inhibitors of
 CYP3A4 (Appendix A)

 7. No chronic anticoagulation

 8. No chronic use of steroids

 9. Karnofsky Performance Scale ≥90% (Appendix C)

 10. Able to provide written, informed consent

 11. For women of child-bearing potential (WOCBP), a negative urine pregnancy test must be
 documented within 7 days prior to the first study intervention

 12. No history of food intolerance to broccoli or pineapple and lime juices

 13. Willing to avoid cruciferous vegetables during the study interventions (Appendix D)

 14. Willing to avoid grapefruit or grapefruit juice 48 hours prior to or during the study

 15. Willing to avoid daily vitamins and anti-inflammatory medications prior to and during
 the study

 16. Potential effects of Broccoli sprout extract on the developing human fetus: The
 effects of Broccoli Sprout Extract on the developing human fetus are unknown. For this
 reason, women of child-bearing potential and men must agree to use adequate
 contraception (hormonal or barrier method of birth control; abstinence) prior to study
 entry and for the duration of study participation. Should a woman become pregnant or
 suspect she is pregnant while she or her partner is participating in this study, she
 should inform her treating physician immediately. Men treated or enrolled on this
 protocol must also agree to use adequate contraception prior to the study and for the
 duration of study participation

 Exclusion Criteria:

 Failure to meet the above criteria