Introduction

Sarcopenia is a progressive, systemic skeletal muscle disease associated with aging that severely affects the quality of life and overall health of older adults. Emerging research demonstrates a strong association between the onset of sarcopenia and gut microbiota.

Aims & Methods

16 out of 30 sarcopenia patients who showed significant skeletal muscle improvement after fecal microbiota transplantation (FMT) were selected and validated by metagenomic, cultured and animal experiments.

Results

Our clinical observations show a marked improvement in sarcopenia symptoms among older patients post FMT, leading to a notable rise in muscle mass (P < 0.05). To delve deeper into this occurrence, we conducted metagenomic sequencing of patient fecal samples pre- and post-treatment. FMT distinctly modified the intestinal microbiota composition. Utilizing germ-free animal models and culturomics, we identified Roseburia inulinivorans as a potentially beneficial strain for the alleviation of sarcopenia. Further experimental validation displayed that Roseburia inulinivorans escalated MyoG and MyoD expression (P < 0.05), augmented muscle mass (P < 0.01) and grip strength (P < 0.01) in mice by boosting branched-chain amino acid metabolism within the host, diminishing sarcopenia symptoms in the process.

Conclusion

These outcomes not only validate the intestinal microbiota-sarcopenia relationship but also introduce novel insights and strategies for sarcopenia management.

References

[1] Tournadre A, Vial G, Capel F, Soubrier M, Boirie Y. Sarcopenia. Joint Bone Spine. 2019. 86(3): 309-314.
[2] DeJong EN, Surette MG, Bowdish D. The Gut Microbiota and Unhealthy Aging: Disentangling Cause from Consequence. Cell Host Microbe. 2020. 28(2): 180-189.
[3] Wang T. Searching for the link between inflammaging and sarcopenia. Ageing Res Rev. 2022. 77: 101611.
[4] Hanna BS, Wang G, Galván-Peña S, et al. The gut microbiota promotes distal tissue regeneration via RORγ+ regulatory T cell emissaries. Immunity. 2023. 56(4): 829-846.e8.
[5] Chen YM, Wei L, Chiu YS, et al. Lactobacillus plantarum TWK10 Supplementation Improves Exercise Performance and Increases Muscle Mass in Mice. Nutrients. 2016. 8(4): 205.
[6] Lee MC, Hsu YJ, Ho HH, et al. Lactobacillus salivarius Subspecies salicinius SA-03 is a New Probiotic Capable of Enhancing Exercise Performance and Decreasing Fatigue. Microorganisms. 2020. 8(4): 545.
[7] Liu X, Wu J, Tang J, et al. Prevotella copri alleviates sarcopenia via attenuating muscle mass loss and function decline. J Cachexia Sarcopenia Muscle. 2023. 14(5): 2275-2288.
[8] Chen LH, Chang SS, Chang HY, et al. Probiotic supplementation attenuates age-related sarcopenia via the gut-muscle axis in SAMP8 mice. J Cachexia Sarcopenia Muscle. 2022. 13(1): 515-531.

Introduction

The pathogenicity of Blastocystis spp. is still debated. Guidelines for donor feces screening differ in their advice to screen for Blastocystis spp., but when tested, its presence is a common reason for exclusion.¹Blastocystis spp. are correlated to increased bacterial alpha-diversity² and distinct bacterial groups³ and therefore its presence may indicate favorable efficacy of fecal microbiota transplantation (FMT). The latest European consensus report no longer advices to reject feces donors that test positive for Blastocystis spp.⁴ Only one paper has been published on human transmission of Blastocystis spp. via frozen FMT.⁵

Aims & Methods

We aimed to investigate the long-term effects of the application of Blastocystis-positive FMT, prepared with fresh feces.
In a FMT trial on FMT with refractory Irritable Bowel Syndrome (IBS) adolescents (age 16 – 20 years) received two administrations (at baseline and after 6 weeks) of fresh allogeneic FMT via nasoduodenal tube prepared with fecal samples from a Blastocystis spp. positive feces donor (NCT03074227). Blastocystis spp. presence and subtyping were determined using microscopy, PCR and Sanger sequencing.

Results

Three recipients received FMT from one donor colonized with Blastocystis subtype 3 (ST3). Two recipients were negative for Blastocystis spp. and one recipient carried ST2 at baseline prior to FMT. After FMT, the two prior-negative recipients tested positive for identical ST3 as the donor after FMT at 12 weeks, but lost carriership of this subtype at week 24 and week 48. The ST2 colonized recipient remained colonized with ST2 and did not acquire ST3 after FMT. Transient adverse events did occur, but did not differ from patients treated with Blastocystis spp.-negative FMT. No FMT-related serious adverse events occurred.

	Recipient	Baseline	Before FMT 1	Donor FMT 1	Before FMT 2	Donor FMT 2
		Week -2	Week 0	Week 0	Week 6	Week 6	Week 12	Week 24	Week 48
1	Woman, 18 y	negative	negative	ST3	ST 3	+, no seq	ST3	negative	negative
2	Woman, 20 y	negative	negative	ST3	+, no seq	ST3	ST 3	ST3	ST3
3	Woman, 16 y	ST2	ST2	ST3	+, no seq	ST3	ST2	ST2	ST2

Table 1: Sanger PCR sequencing results of frozen feces samples from three adolescents with IBS and one healthy donor. Samples were collected before, during and after the FMT treatment schedule with follow-up unto week 48. Light blue cells indicate a different sequence order of Blastocystis ST3 than the Blastocystis ST3 indicated in the dark blue cells. Grey cells indicate columns representing donor samples.
Abbreviations: FMT; fecal microbiota transplantation, ST; subtype, seq; sequence, y; years.

Conclusion

We present the first long-term results of Blastocystis spp. transmission by fresh FMT in three cases. Transient colonization with Blastocystis spp. did occur without the occurrence of FMT-related serious adverse events.

References

1. Benard MV, de Bruijn CMA, Fenneman AC, Wortelboer K, Zeevenhoven J, Rethans B, et al. Challenges and costs of donor screening for fecal microbiota transplantations. PLoS One. 2022;17(10):e0276323.
2. Tito RY, Chaffron S, Caenepeel C, Lima-Mendez G, Wang J, Vieira-Silva S, et al. Population-level analysis of Blastocystis subtype prevalence and variation in the human gut microbiota. Gut. 2019;68(7):1180-9.
3. Nieves-Ramirez ME, Partida-Rodriguez O, Laforest-Lapointe I, Reynolds LA, Brown EM, Valdez-Salazar A, et al. Asymptomatic Intestinal Colonization with Protist Blastocystis Is Strongly Associated with Distinct Microbiome Ecological Patterns. mSystems. 2018;3(3).
4. Keller JJ, Ooijevaar RE, Hvas CL, Terveer EM, Lieberknecht SC, Hogenauer C, et al. A standardised model for stool banking for faecal microbiota transplantation: a consensus report from a multidisciplinary UEG working group. United European Gastroenterol J. 2021;9(2):229-47.
5. Terveer EM, van Gool T, Ooijevaar RE, Sanders I, Boeije-Koppenol E, Keller JJ, et al. Human Transmission of Blastocystis by Fecal Microbiota Transplantation Without Development of Gastrointestinal Symptoms in Recipients. Clin Infect Dis. 2020;71(10):2630-6.

Disclosure

CYP has received research grants from Gilead and Perspectum, consultancy fee from Chemomab and NGM, and speaker’s fee from Tillotts. The other authors disclose no conflicts.

Introduction

Fibrostenosis represents one of the major clinical complications of Crohn’s Disease (CD). However, the molecular mechanisms underlying intestinal fibrogenesis remain poorly understood and anti-fibrotic therapy has not yet been developed. Growing evidence supports that oxidative stress and the epithelial-mesenchymal transition (EMT) process contribute to the development and progression of fibrosis.

Aims & Methods

Based on these evidences, we assessed the role of lipids in fibrogenesis and whether mitochondria dysfunction can support fibrosis formation.
Spatial metabolomics analysis and adipophilin (ADFP) staining were performed on surgical specimens from fibrotic ileum and the surrounding region (healthy) derived from CD patients. Fibroblasts isolated from fibrotic and healthy ileum and human induced pluripotent stem cell-derived intestinal organoids (iHOs) were treated for 1-3 weeks with lipid mix (2%) and analyzed by RNA sequencing. Pathways significantly modulated were validated by RT-PCR, WB, FACS analysis, and proliferative, migratory and contraction assays, and mitochondrial functions by Seahorse analyzer.

Results

Spatial metabolomics revealed a significant accumulation of lipids characterized by very long-chain fatty acids in the fibrotic area compared to surrounding healthy regions, which was further confirmed by ADFP staining. RNA-seq analysis showed overlapping dysregulated pathways between healthy fibroblasts stimulated with lipids and fibrotic fibroblasts, in particular genes related to cell cycle, cytoskeletal reorganization, migration, contractility and cell metabolism (oxidative stress) which were further confirmed by functional assays. Among the genes related to oxidative stress, lipid stimulation significantly decreased NFE2L2 and promoted an increase in CPT1A, gene encoding for a fatty oxidation mediator.
Moreover, lipid-stimulated fibroblasts showed a reduction in mitochondrial respiration, particularly basal and maximal respiration, and spare respiratory capacity by Seahorse analysis. On the other hand, long exposure of iHOs to lipids promoted the EMT process through the upregulation of TWIST, ZEB, and VIM.

Conclusion

Lipid stimulation induced myofibroblast activation through oxidative stress and alteration of mitochondrial metabolism, and increased the expression of EMT transcription factors in intestinal epithelial cells (iHOs) that may support fibroblast-mediated pro-fibrotic activity. These data indicate that regulators of fatty acid oxidation involved in the fibrotic process may represent new potential targets for the treatment of fibrostenotic CD patients.

Introduction

Since 2006 at our hospital, lifestyle modification, such as diet and exercise, for obese patients has been provided through a team approach with a physician, dietician, nurse, physical therapist and psychologist. From 2016, a surgeon and anesthesiologist were added to the team for metabolic/bariatric surgery (laparoscopic sleeve gastrectomy: LSG).

Aims & Methods

The aim of this study is to clarify the characteristics of obese patients with diabetes mellitus (DM) for whom LSG was effective according to body mass index (BMI). Of 24 diabetic patients who were received LSG between January 2017 and December 2021 with follow-up period of more than 1 year, 4 were with a BMI of < 35, 7 were with a BMI of 35 - 40, and 13 were with a BMI of 40 or more. The mean age was 51, 49 and 45 years. respectively. Complete remission of postoperative DM was defined as HbA1c < 6% without drug therapy, and incomplete remission was defined as a decrease in the amount of drug therapy among the other patients.

Results

Preoperative HbA1c (%) was 8.6, 7.0 and 8.0, and preoperative serum c-peptide (CPR) (ng/ml) was 1.9, 3.1 and 3.1, and preoperative ABCD score was 2.0, 4.4 and 6.0, respectively. There were no differences of preoperative complications, preoperative weight loss rate, postoperative weight loss rate, and postoperative body composition change rate. The postoperative complete DM remission rate (%) was 25, 71 and 69, respectively, and the drug dose was reduced in all patients, even in those with incomplete remission.

Conclusion

Regardless of BMI, complete remission of DM was achieved by performing LSG when CPR was maintained, even in patients with poorly controlled DM. On the other hand, even in patients with decreased CPR, benefits such as postoperative weight loss and drug reduction were observed. LSG should be actively considered in obese patients with DM.

Introduction

Acute Severe Ulcerative Colitis (ASUC) is a serious, potentially life-threatening condition. Hospital admission and prompt medical therapy with intravenous steroids is crucial, but rescue medical therapy with infliximab is frequently needed. Despite its frequency and relevance, there is no clear evidence about the optimal infliximab (IFX) dosing and timing in this context.

Aims & Methods

In a prospective multicentre, international cohort study, consecutive patients hospitalized for ASUC between 2/2019-1/2020 were included. The primary outcome was surgery during the first 90 days after admission (failure of medical treatment). For this analysis we identified and analyzed all patients receiving IFX as rescue therapy during the index admission. We used descriptive statistics along with non-parametric tests to determine patient characteristics and outcomes between IFX usage patterns.

Results

A total of 823 patients from 123 centers in 32 countries in five continents were included (median age 39 years [IQR, 28-54], 56% male, 64% non-smokers). IFX was started in 159 patients (19%) after a median of 6 days (IQR, 4-8) after starting steroids, with 27% after >7 days. The initial IFX dose was mostly 5 mg/kg (74%), followed by 10 mg/kg (23%). The overall response was considered adequate in 74%, this was numerically superior among those receiving 10 mg/kg (83% vs 74%, p=0.5). In 41 patients a second dose of IFX (26%) was administered during the admission. After a first dose of IFX 5 mg/kg or 10 mg/kg, most patients received a subsequent infusion with the same dosing (92% and 79%, respectively). Colectomy was performed in 35 patients (22%), without statistically significant differences between those with an initial dose of 5 or 10 mg/kg (23% vs 19%, p=0.6).

Conclusion

IFX is a relevant rescue therapy, required in around one in five patients with ASUC. Most patients receive 5 mg/kg, and this is usually followed by the same dosage. Still a proportion of patients receive higher doses with 10 mg/kg, but colectomy risk remains similar between both groups.

Introduction

The therapeutic goal in inflammatory bowel disease (IBD), which includes Crohn´s disease (CD) and ulcerative colitis (UC), is the control of the inflammatory process through drugs (biologic and JAK-inhibitor therapies) that generally have a high cost and may have side effects. Unfortunately, only one-third of patients respond to these treatments and there are no markers that accurately predict this response.

Aims & Methods

The aim of this study was to identify potential serum metabolic biomarkers predictive of response to different drugs for IBD at baseline (before treatment), which could facilitate the selection of the most appropriate therapy for each patient. For this purpose, a prospective and multicenter study was conducted including CD and UC patients who initiate biologic (anti-TNF, ustekinumab or vedolizumab) or JAK-inhibitor (tofacitinib) therapies. Patients were assessed at 14 weeks to determine if they had achieved endoscopic response, based on the SES-CD and on the endoscopic sub score of the Partial Mayo Score. The analysis of the serum metabolome and deconvolution of the serum lipoprotein profile into 110 fractions were conducted using nuclear magnetic resonance for quantitative differential analysis comparing baseline serum samples of responders compared with non-responders.

Results

A total of 150 patients were included (30 CD anti-TNF, 30 UC anti-TNF, 30 CD ustekinumab, 30 UC vedolizumab, and 30 UC tofacitinib). Table 1 shows a summary of the most relevant results obtained during the metabolomic analysis, which revealed metabolites and lipoproteins (common or drug-specific) with significantly altered levels (p<0.05) in the baseline serum samples of subsequent responders compared to non-responders. Moreover, based on ROC curves, potential biomarkers with good sensitivity and specificity for distinguishing between responders and non-responders before treatment were identified. Finally, our analysis identified alanine, aspartate, glutamate, arginine and butanoate metabolism, and the citrate cycle, as the main baseline altered metabolic pathways that could be associated with the response to IBD treatment.

Table 1. Summary of identified potential metabolic biomarkers predictive of endoscopic response to IBD treatment.

Study group	Number of Biomarkers*		Best AUC values**
	Metabolites	Lipoproteins
CD anti-TNF	2	3	0.83; 0.81; 0.78; 0.75; 0.74
UC anti-TNF	1	7	0.88; 0.84; 0.83; 0.77; 0.76; 0.73
CD ustekinumab	4	24	0.96; 0.95; 0.94; 0.91; 0.90; 0.89; 0.87; 0.80
UC vedolizumab	5	8	0.90; 0.79; 0.78
UC tofacitinib	3	8	0.98; 0.96; 0.91; 0.88; 0.87; 0.85

​*Biomarkers with significantly altered levels (p<0.05) in the baseline serum of responders vs non-responders patients. **AUC corresponding to the biomarkers with the best sensitivity and specificity for distinguishing between responders and non-responders patients before treatment. AUC: Area Under the Curve calculated using the Metabolanalyst software in univariate analysis; CD: Crohn's disease; IBD: Inflammatory bowel disease; UC: Ulcerative colitis.

Conclusion

The present study represents an effort towards personalized medicine in targeted therapy selection in IBD patients, which is an important unmet need. Although it needs further validation, we were able to identify a number of serum metabolites and lipoproteins with the potential of predicting response to biologic and JAK-inhibitor therapies.