Overview of the study:
This is a four-week randomised double-blind parallel-arm food supplement intervention study that will determine the effect of fructose on metabolism in people with MASH, compared to people with simple steatosis, or controls without liver disease. The study will involve consent (visit 0A) and screening (visit 0B), followed by two study stages (see Figure 1), where participants will undergo study visits and procedures as detailed in Table 1. Each participant will attend for a total of up to 5 study visits: consent (visit 0A) and screening (visit 0B); Baseline (visit 1); pre-intervention (visit 2); post-intervention (visit 3). Consent (visit 0A) and screening (visit 0B) visits may be combined into one visit if feasible and the participant would like to do so. Otherwise, visits will be conducted on separate days as close together as practical for the participant and research team (ideally no more than 3-4 weeks). Visits 1 to 3 will each last approximately 8 hours, at which participants will consume a test meal and have hourly blood samples taken via a cannula for approximately 6h.
A subset of participants (maximum of n=24; n=12 MASH; n=12 steatosis/control) will be invited to take part in an optional sub-study. This will involve the same study visits and procedures as the main study but will additionally measure how glucose or fructose is metabolised in the postprandial state by addition of a small amount of glucose/fructose stable isotope tracer (13C6-fructose or 13C6-glucose) to the metabolic test meal. Participants will also provide breath samples at hourly intervals during the 6h feeding period.
There will be no change to routine patient care for MASH and MASLD participants during the study period. Any clinically significant results emerging from study participation will be shared with participants' GP or care team.
Description of the study intervention:
Following consent (visit 0A) and screening (visit 0B), participants will receive a study guide with details of the main study visits and procedures, and instructions on how to complete the 3-day food diaries. At the baseline visit (visit 1), participants will receive dietary counselling and other information to help them follow a diet low in free sugars, including fructose, glucose, and sucrose. The nutrition researcher and participant will agree on a personalised list of suggested low-sugar meals, drinks and snacks that are suitable to be consumed during the study. Participants will be given a choice of food, drink and snack items to be delivered each two weeks as the basis for a low-sugar diet. The nutrition researcher will offer ongoing support to ensure compliance via regular phone calls, messages, and at study visits as needed.
Following the Baseline visit (visit 1), participants will undergo a two-week run-in period on the low-sugar diet, after which they will be randomised to either group A or B for the supplementation period. Both groups will be maintained on the low-sugar diet and additionally receive supplemental 100g of fructose (group A) or glucose (group B) per day for a period of 14 days. All sachets will be labelled as 'Sugar Supplement' or similar.
Each 100g daily dose of glucose or fructose will be prepared by an independent researcher at King's College London and divided into two labelled sachets (50g each). Participants will be directed to dissolve each of the sugar sachets in approximately 300 ml of water and to consume these solutions in between their primary meals. All study participants will be provided with a daily multivitamin supplement to prevent any risk of nutrient deficiencies due to the low fruit content of the prescribed diet.
Dietary intake will be analysed through a series of 3-day food diaries completed contemporaneously prior to each of the study visits (baseline, pre- and post-intervention). The UK Composition of Foods Integrated Dataset will be used to compute the macro- and micronutrient composition, as well as the caloric value, of the diet. The dietary intervention will be compiled by an expert nutrition researcher and overseen by a qualified dietitian.
Participants will attend five study visits, including consent (visit 0A) and screening (visit 0B), a baseline visit (visit 1; before starting the low-sugar diet), as well as visits pre-intervention (visit 2) and post-intervention (visit 3), as detailed in section 7.3. Consent (visit 0A) and screening (visit 0B) visits may be combined into one visit if feasible and the participant would like to do so.
Participants will be remunerated for their time and commitment to completing the study procedures.
Visits for the study:
Pre-screening: Potential participants will be recruited from liver clinics (MASH and steatotic patients) or via the PIC site (King's College Hospital); control (healthy) participants will be recruited via email or poster adverts. All potential participants will receive a copy of the Participant Information Sheet and be given time to read and consider this. Prior to attending for a consent or screening visit, potential participants will undergo a pre-screening call or discussion with either a member of their clinical care team (MASH/steatosis patients) or a member of the study research team (controls) to check their eligibility against the inclusion/exclusion criteria prior to attending in person.
Consent (Visit 0A): Prior to the consent visit, participants will receive the participant information sheet and have the opportunity to consider this (and discuss it with others if they wish). Participants will attend for the visit and have the opportunity to ask questions or discuss the study with a member of the research team, before offering their informed consent. Participants will receive a copy of their signed consent form and agree on a suitable date for the screening visit.
Screening (visit 0B): Participants will answer questions concerning medical history, have anthropometrics and vital signs measured by a researcher, and (if required to confirm eligibility) have a Fibroscan and fasted blood sample taken. Participants will be informed of their eligibility following the visit and be provided with a copy of the study guide, which includes instructions on how to complete the 3-day food diaries.
Visits 1, 2 and 3: Participants will be sent a reminder to complete each of the 3-day food diaries prior to each visit. They will also be given a low-fat standardised meal to consume the night before and will attend in the morning (at approximately 8am) after an overnight fast (≥12h). Participants will provide fresh stool, urine, and saliva samples, and researchers will take anthropometric measurements and vital signs (e.g. blood pressure). Measurements will be conducted utilising Vibration-Controlled Transient Elastography with Controlled Attenuation Parameter (VCTE CAP).
Participants will then undergo a 6-hour post-prandial feeding protocol, which will measure the metabolic response to consuming a standardised mixed nutrient test meal. Participants will be cannulated and have fasted blood samples taken, before consuming the mixed-nutrient meal. Blood samples will be taken every hour over the subsequent 6-hour period to test metabolic response to the test meal, including markers of lipid and glucose metabolism. Participants will not be able to eat anything else during this period but will be offered ad libitum drinking water.
In the optional sub-study, participants will additionally consume 13C6-glucose or 13C6-fructose stable isotope tracer as part of the test meal and provide breath samples at hourly intervals, in addition to the hourly blood samples. This will allow the metabolism of glucose and fructose to be measured in blood (13C-palmitate for de novo lipogenesis) and breath (13CO2 for glucose/fructose oxidation) samples to provide additional insight into how the sugars are metabolised.
Following the post-prandial procedure, the cannula will be removed. Participants will be offered something to eat, given any additional instructions, dietary counselling, or study supplies required for the next stage of the study and will then be able to leave.
