Fasting-mimicking Diet Research Articles

The Role of Calorie Restriction in Cancer Prevention, Cancer Treatment, Longevity, and in Reducing Cellular Stress

Cancer is one of the top main illnesses worldwide. The aging of the population and the widespread presence of risk factors including obesity, smoking, and/or bad eating habits are expected to contribute to its further rise. Chemotherapy, radiation therapy, and surgery continue to be the cornerstones of cancer treatment, despite the gradual replacement or supplementation of these methods by innovative treatments. The American Cancer Society suggests a high-calorie, high-protein diet for cancer patients undergoing chemotherapy. Furthermore, there are no established recommendations for the kind of diet that would significantly reduce cancer rates. However, calorie restriction (CR) and fasting, two common methods of reducing caloric consumption, show a broad variety of positive benefits that may prevent malignancies and boost the effectiveness of cancer treatments. Periodic fasting (PF), fasting-mimicking diets, and restricted food intake without first a decrease in calories have become popular as interventions with the potential to be widely used to prevent and treat cancer, while chronic CR provides both beneficial and detrimental effects and major compliance challenges. In this article, we discuss the impact that calorie restriction has in preventing cancer, aiding in cancer treatment, producing cellular protection and chemotherapy resistance in animal models and early human investigations.

Fasting mimicking diet inhibits tumor-associated macrophage survival and pro-tumor function in hypoxia: implications for combination therapy with anti-angiogenic agent

BackgroundRecent research shows that tumor-associated macrophages (TAMs) are the primary consumers of glucose in tumor tissue, surpassing that of tumor cells. Our previous studies revealed that inhibiting glucose uptake impairs the survival and tumor-promoting function of hypoxic TAMs, suggesting that glucose reduction by energy restriction (calorie restriction or short-term fasting) may has a significant impact on TAMs. The purpose of this study is to verify the effect of fasting-mimicking diet (FMD) on TAMs, and to determine whether FMD synergizes with anti-angiogenic drug apatinib via TAMs.MethodsThe effect of FMD on TAMs and its synergistic effects with apatinib were observed using an orthotopic mouse breast cancer model. An in vitro cell model, utilizing M2 macrophages derived from THP-1 cell line, was intended to assess the effects of low glucose on TAMs under hypoxic and normoxic conditions. Bioinformatics was used to screen for potential mechanisms of action, which were then validated both in vivo and in vitro.ResultsFMD significantly inhibit the pro-tumor function of TAMs in vivo and in vitro, with the inhibitory effect being more pronounced under hypoxic conditions. Additionally, the combination of FMD-mediated TAMs inhibition with apatinib results in synergistic anti-tumor activity. This effect is partially mediated by the downregulation of CCL8 expression and secretion by the mTOR-HIF-1α signaling pathway.ConclusionsThese results support further clinical combination studies of FMD and anti-angiogenic therapy as potential anti-tumor strategies.Graphical

248P Targeting triple-negative breast cancer metabolism with neoadjuvant chemotherapy plus fasting-mimicking diet plus/minus metformin: The BREAKFAST trial

248P Targeting triple-negative breast cancer metabolism with neoadjuvant chemotherapy plus fasting-mimicking diet plus/minus metformin: The BREAKFAST trial

Fasting-mimicking diet alleviates inflammatory pain by inhibiting neutrophil extracellular traps formation and neuroinflammation in the spinal cord

BackgroundNeutrophil extracellular traps (NETs) promote neuroinflammation and, thus, central nervous system (CNS) disease progression. However, it remains unclear whether CNS-associated NETs affect pain outcomes. A fasting-mimicking diet (FMD) alleviates neurological disorders by attenuating neuroinflammation and promoting nerve regeneration. Hence, in this study, we explore the role of NETs in the CNS during acute pain and investigate the role of FMD in inhibiting NETs and relieving pain.MethodsThe inflammatory pain model was established by injecting complete Freund’s adjuvant (CFA) into the hind paw of mice. The FMD diet regimen was performed during the perioperative period. PAD4 siRNA or CI-amidine (PAD4 inhibitor) was used to inhibit the formation of NETs. Monoamine oxidase-B (MAO-B) knockdown occurred by AAV-GFAP-shRNA or AAV-hSyn-shRNA or was inhibited by selegiline (an MAO-B inhibitor). The changes in NETs, neuroinflammation, and related signaling pathways were examined by western blot, immunofluorescence, ELISA, and flow cytometry.ResultsIn the acute phase of inflammatory pain, NETs accumulate in the spinal cords of mice. This is associated with exacerbated neuroinflammation. Meanwhile, inhibition of NETs formation alleviates allodynia and neuroinflammation in CFA mice. FMD inhibits NETs production and alleviates inflammatory pain, which is enhanced by treatment with the NETs inhibitor CI-amidine, and reversed by treatment with the NETs inducer phorbol 12-myristate 13-acetate (PMA). Mechanistically, the neutrophil-recruiting pathway MAO-B/5-hydroxyindoleacetic acid (5-HIAA) / G-protein-coupled receptor 35 (GPR35) and NETs-inducing pathway MAO-B/ Reactive oxygen species (ROS) are significantly upregulated during the development of inflammatory pain. MAO-B is largely expressed in astrocytes and neurons in the spinal cords of CFA mice. However, knockdown or inhibition of MAO-B effectively attenuates CFA-induced inflammatory pain, NETs formation, and neuroinflammation in the spinal cord. Moreover, within rescue experiments, MAO-B inhibitors synergistically enhance FMD-induced pain relief, NETs inhibition, and neuroinflammation attenuation, whereas supplementation with MAO-B downstream molecules (i.e., 5-HIAA and PMA) abolished this effect.ConclusionsNeutrophil-released NETs in the spinal cord contribute to pain development. FMD inhibits NETs formation and NETs-induced neuroinflammation by inhibiting the MAO-B/5-HIAA/GPR35 and MAO-B/ROS pathways in astrocytes and neurons, thereby relieving pain progression.BHScZMHeZam_oB9V3iTexJVideo

Fasting mimicking diet in mice delays cancer growth and reduces immunotherapy-associated cardiovascular and systemic side effects

Immune checkpoint inhibitors cause side effects ranging from autoimmune endocrine disorders to severe cardiotoxicity. Periodic Fasting mimicking diet (FMD) cycles are emerging as promising enhancers of a wide range of cancer therapies including immunotherapy. Here, either FMD cycles alone or in combination with anti-OX40/anti-PD-L1 are much more effective than immune checkpoint inhibitors alone in delaying melanoma growth in mice. FMD cycles in combination with anti-OX40/anti-PD-L1 also show a trend for increased effects against a lung cancer model. As importantly, the cardiac fibrosis, necrosis and hypertrophy caused by immune checkpoint inhibitors are prevented/reversed by FMD treatment in both cancer models whereas immune infiltration of CD3+ and CD8+ cells in myocardial tissues and systemic and myocardial markers of oxidative stress and inflammation are reduced. These results indicate that FMD cycles in combination with immunotherapy can delay cancer growth while reducing side effects including cardiotoxicity.

A Biodegradable Nucleotide Coordination Polymer for Enhanced NSCLC Therapy in Combination with Metabolic Modulation.

Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) in the treatment of non-small cell lung cancer (NSCLC) still face challenges of acquired resistance and non-negligible side effects. To overcome these limitations, a biodegradable coordination polymer using guanine deoxynucleotide and ferrous iron (dGNP) is developed for targeted delivery of EGFR-TKIs. dGNPs can efficiently target nucleoside transporters in tumor cells that are regulated by fasting-mimicking diet (FMD). Meanwhile, FMD can augment the therapeutic efficacy of EGFR-TKIs by suppressing EGFR tyrosine kinase phosphorylation and related downstream pathways. In vivo results demonstrate that EGFR-TKIs-laden dGNPs combined with FMD treatment exhibit superior antitumor efficacy and reduced side effect. This study provides an innovative approach to enhance the therapeutic efficacy of EGFR-TKIs through nucleotide nanocarrier and metabolic modulation.

Adding fasting-mimicking diet to first-line carboplatin-based chemotherapy is associated with better overall survival in advanced triple-negative breast cancer patients: A subanalysis of the NCT03340935 trial.

Severe calorie restriction, in the form of cyclic fasting or fasting-mimicking diets (FMDs), boosts the antitumor activity of cytotoxic chemotherapy in mouse models of triple-negative breast cancer (TNBC). This effect is mostly mediated by fasting/FMD-induced reduction of plasma glucose concentration and by a boost in antitumor immunity. However, clinical evidence that cyclic FMD may impact on the outcomes of advanced TNBC (aTNBC) patients is lacking. We compared the overall survival (OS) of 14 aTNBC patients receiving first-line carboplatin-gemcitabine plus cyclic FMD in the context of the NCT03340935 trial with the OS of 76 consecutive aTNBC patients treated with carboplatin-based chemotherapy alone at Fondazione IRCCS Istituto Nazionale dei Tumori. Multivariable Cox regression models were used to adjust the prognostic impact of FMD for other prognostic variables. Patients undergoing cyclic FMD in combination with carboplatin-gemcitabine had better OS when compared to patients receiving chemotherapy alone (median OS 30.3 months, 95% CI 18-NR, vs 17.2 months, 95% CI 15.3-25.1, log-rank P value .041). Multivariable analysis confirmed an association between FMD use and better OS (HR: 0.40; 95% CI: 0.19-0.86; P = .019) also after propensity score-based matching according to patient ECOG PS and the presence of de novo metastatic disease (HR: 0.41; 95% CI: 0.21-0.83; P = .013). Cyclic FMD in combination with first-line chemotherapy may improve clinical outcomes in aTNBC patients. Our study paves the way for conducting phase II trials to investigate if cyclic FMD can increase the antitumor activity/efficacy of chemotherapy or chemoimmunotherapy in patients with early-stage TNBC or aTNBC.

Fasting-mimicking diet drives antitumor immunity against colorectal cancer by reducing IgA-producing cells.

As a safe, feasible, and inexpensive dietary intervention, fasting-mimicking diet (FMD) exhibits excellent antitumor efficacy by regulating metabolism and boosting antitumor immunity. A better understanding of the specific mechanisms underlying the immunoregulatory functions of FMD could help improve and expand the clinical application of FMD-mediated immunotherapeutic strategies. In this study, we aimed to elucidate the role of metabolic reprogramming induced by FMD in activation of antitumor immunity against colorectal cancer (CRC). Single-cell RNA sequencing (scRNA-seq) analysis of intratumoral immune cells revealed that tumor-infiltrating IgA+ B cells were significantly reduced by FMD treatment, leading to the activation of antitumor immunity and tumor regression in murine CRC models. Mechanistically, FMD delayed tumor growth by repressing B cell class switching to IgA. Therefore, FMD-induced reduction of IgA+ B cells overcame the suppression of CD8+ T cells. The immunoregulatory and antitumor effects of FMD intervention were reversed by IgA+ B cell transfer. Moreover, FMD boosted fatty acid oxidation (FAO) to trigger RUNX3 acetylation, thus inactivating Cα gene transcription and IgA class switching. IgA+ B cell expansion was also impeded in patients placed on FMD, while B cell expression of CPT1A, the rate-limiting enzyme of FAO, was increased. Furthermore, CPT1A expression was negatively correlated with both IgA+ B cells and IgA secretion within CRC. Together, these results highlight that FMD holds great promise for treating CRC. Furthermore, the degree of IgA+ B cell infiltration and FAO-associated metabolic status are potential biomarkers for evaluating FMD efficacy.

Fasting-mimicking diet: a metabolic approach for the treatment of breast cancer.

Metabolic reprogramming is a new and potentially targetable hallmark of cancer. In recent years, fasting and fasting-mimicking diets (FMDs) have been tested as anticancer strategies both in preclinical experiments and in clinical trials. In this review, we aim at summarizing the available evidence about the antitumour activity of these approaches in preclinical breast cancer models, as well as results from clinical trials investigating fasting/FMD in breast cancer patients. Preclinical evidence demonstrated that nutrient deprivation boosts the antitumor activity of chemotherapy, immunotherapy or targeted therapies in triple-negative breast cancer (TNBC) and HR+/HER2 models through both cell-autonomous antitumour effects in cancer cells and favourable modifications in intratumor immune cells. Several clinical experiences demonstrated that fasting/FMD is feasible and well tolerated in combination with standard treatments in BC patients, and that it could reduce chemotherapy-related toxicities. Finally, despite the absence of randomized trials demonstrating the antitumor activity of fasting/FMD in breast cancer patients, preliminary clinical reports suggest that this experimental nutritional strategy may enhance chemotherapy activity. Randomized clinical trials are ongoing to validate these results at a larger scale. Fasting/FMD is a promising therapeutic approach in patients with breast cancer; ongoing and future trials will confirm their role in improving breast cancer care.

Verification of fasting-mimicking diet to assist monotherapy of human cancer-bearing models

The efficacy of a single clinical nanodrug for cancer treatment is still unsatisfactory, especially for drug-resistant cancer. Herein, we applied a fasting-mimicking diet (FMD) approach via dietary intervention to assist single clinical nanodrug for breast or ovarian cancer treatments instead of using multi-drug therapies which might cause adverse side effects. Specifically, we adopted Doxil or Abraxane to treat human breast tumor-bearing nude mice and Doxil to treat the human ovarian tumor and drug-resistant ovarian tumor-bearing nude mice under FMD conditions, respectively. According to the results, the FMD condition can promote the cellular uptake and cytotoxicity of a single nanodrug, reduce the ATP level in drug-resistant tumor cells to hinder drug efflux, normalize tumor blood vessels, relieve tumor hypoxia, and increase the accumulation of nanodrugs at tumor sites, thereby enhancing the therapeutic effects on these types of human cancers. Collectively, these results demonstrate that the FMD strategy of significance can become a practical, alternative, and promising assistant for single nanodrug for enhancing cancer therapy and clinical translation.

Scientific evidence of foods that improve the lifespan and healthspan of different organisms.

Age is a risk factor for numerous diseases. Although the development of modern medicine has greatly extended the human lifespan, the duration of relatively healthy old age, or 'healthspan', has not increased. Targeting the detrimental processes that can occur before the onset of age-related diseases can greatly improve health and lifespan. Healthspan is significantly affected by what, when and how much one eats. Dietary restriction, including calorie restriction, fasting or fasting-mimicking diets, to extend both lifespan and healthspan has recently attracted much attention. However, direct scientific evidence that consuming specific foods extends the lifespan and healthspan seems lacking. Here, we synthesized the results of recent studies on the lifespan and healthspan extension properties of foods and their phytochemicals in various organisms to confirm how far the scientific research on the effect of food on the lifespan has reached.

The Potential of Fasting-Mimicking Diet as a Preventive and Curative Strategy for Alzheimer's Disease.

This review examines the potential of fasting-mimicking diets (FMDs) in preventing and treating Alzheimer's disease (AD). FMDs are low-calorie diets that mimic the physiological and metabolic effects of fasting, including the activation of cellular stress response pathways and autophagy. Recent studies have shown that FMDs can reduce amyloid-beta accumulation, tau phosphorylation, and inflammation, as well as improve cognitive function in animal models of AD. Human studies have also reported improvements in AD biomarkers, cognitive functions, and subjective well-being measures following FMDs. However, the optimal duration and frequency of FMDs and their long-term safety and efficacy remain to be determined. Despite these uncertainties, FMDs hold promise as a non-pharmacological approach to AD prevention and treatment, and further research in this area is warranted.

419-P: Glucose-Induced Cellular Resistance to Methylglyoxal Is Lost in Patients with Diabetes Complications and Further Deteriorated after Caloric Restriction

Aim: Glucose stress can be protective against dicarbonyl-mediated toxicity. This study aimed to investigate cellular resistance against methylglyoxal (MG) after glucose stress in healthy humans (CON), patients with (T2D+) and without diabetes complications (T2D-) and whether this is affected by caloric restriction. Fibroblast growth factor 21 (FGF21) and β- hydroxybutyrate (β-HB) were analyzed as marker for metabolic adaptation to stress response and ketogenesis. Methods: Peripheral blood mononuclear cells were isolated before and 2h after oGTT and incubated at each time-point with increasing MG-levels for 24hrs. Cell viability was measured with CellTiter-Glo to determine changes of MG-resistance at 2h after oGTT (ΔEC50). Experiments were repeated after a 5-day fasting-mimicking diet (FMD). FGF21 and β-HB were measured in venous blood. Results: Thirty participants (18m/12f, age 64.6 [62.2, 67.0] y, BMI 28.6 [27.2, 30.0] kg/m2) were included. HbA1c was higher in T2D+ (7.4 [6.9, 7.9] %) and T2D- (7.0 [6.2, 7.7] %) compared to CON (5.5 [5.1, 5.8] %, p<0.001), age and BMI were comparable. ΔEC50 of MG increased in CON before and after FMD (12.9% vs. 12.6%), was unaffected in T2D- (1.8% vs. -1.7%) and decreased in T2D+ after FMD (-3.9% vs. -19.0%, p<0.01) and when compared to T2D- and CON (p<0.05). ΔFGF21 increased in CON after FMD (7.7% vs. 115.0%, p<0.05) and tended to be higher compared to T2D- and T2D+ (115.0% vs. 69.3% vs. 47.5%, p=0.5). ΔFGF21 correlated with β-HB (r = 0.48, p<0.01), with strongest correlation for T2D+. Conclusion: Acute glucose stress induces cellular resistance to MG in healthy humans, a response which is lost in T2D with diabetic complications and aggravates after caloric restriction. Our data suggest for an association between loss of capacity for handling dicarbonyl stress and the presence of diabetic complications. FGF21 might play an important role in the metabolic adaptation to dicarbonyl stress and ketosis. Disclosure E.V.Rauchhaupt: None. C.Rodemer: None. E.Kliemank: None. S.Kopf: Advisory Panel; Novo Nordisk, Bayer Inc., Recordati Rare Disease, Speaker's Bureau; Novo Nordisk, Boehringer-Ingelheim. S.Herzig: Consultant; Novo Nordisk, Actimed, Research Support; Novo Nordisk, Boehringer-Ingelheim, Almac. P.Nawroth: None. J.Szendroedi: None. J.Zemva: None. A.Sulaj: None. Funding DFG; German Research Foundation (236360313-SFB, 1118DZD); German Center for Diabetes Research (82DZD07C2G)

Role of Fasting Mimicking Diet in Farnesoid x Receptor for Suppressing Epithelial-to-Mesenchymal Transition, Cell Cycle Progression, and Viability of Prostate Cancer Cells

The systemic and resistant nature of metastatic castration-resistant prostate cancers (mCRPC) renders it largely incurable even after intensive multimodal therapy. Proliferation, survival, and epithelial-mesenchymal transition (EMT) are three fundamental events that are deeply linked to carcinogenesis. Hence, it is necessary to find a new combination of several therapies, targeting those vital mechanisms without causing side effects. Significant research works have shown differential low expression of the metabolic Farnesoid X receptor (FXR) in primary and metastatic prostate cancer suggesting their importance in prostate pathogenesis. Obticholic acid (INT 747), a potent FXR agonist is widely used in primary biliary cholangitis, and Fasting mimicking Diet (FMD) both were drastically showed effects on different cancer progression. We hypothesized that FXR and FMD may inhibit proliferation and the metastatic phenotype in PC-3 prostate cancer cells. Analyses of the cell viability, cell cycle, migration, and matrigel invasion assays were performed to elucidate how INT 747 and /or FMD functions in prostate cancer. In this study, INT 747 treatment caused apoptotic morphological changes and significantly reduced the survival of PC-3 cells incubated in normal mediums. Furthermore, we showed that the combination of the INT 747 and FMD was much more harmful to cancer cells than the treatment with INT 747 or FMD alone. Moreover, our study showed that INT 747 either alone or combined with FMD robustly induced cell cycle arrest at the S phase. Interestingly, the combination treatment on PC-3 cells not only showed several lines of evidence of apoptotic cells death but also inhibited carcinogenic potential as evaluated by impairment of spheroid formation capacity and delayed wound healing and matrigel invasion. At the cellular level, FXR activation resulted in down-regulation of procaspase -3, vimentin, and MMP9, which triggers apoptotic cell death, cell cycle arrest, and switch from mesenchymal to an epithelial phenotype. Collectively, FXR activation alone markedly decreases, and when combined with FMD abrogates the survival and carcinogenic potential of metastatic prostate cancer cells.

DIRECT2: A randomized phase III trial with a fasting mimicking diet program to improve chemotherapy in patients with hormone receptor positive, HER2-negative breast cancer—A Dutch Breast Cancer Research Group study (BOOG).

TPS630 Background: Preclinical research has shown that fasting and/or a fasting mimicking diet (FMD) in addition to chemotherapy can enhance cancer treatment effectivity and reduce side-effects. The previous DIRECT1 phase 2 study (de Groot S, Nat. Commun. 2020) indicated that FMD could increase effectivity of neoadjuvant chemotherapy in patients with HER2-negative breast cancer. Methods: DIRECT2 is a multicenter, randomized, open label phase 3 study that investigates whether FMD compared to a regular diet during neoadjuvant chemotherapy can improve the pathological and/or radiological response rate in patients with hormone receptor positive (HR+), HER2-negative, stage II/III breast cancer. Secondary endpoints include quality of life, cognition, toxicity ≥grade 3 and the immunomodulating effect of FMD. The FMD consists of a low-caloric diet of 600kcal/day starting 3 days prior to, and on the day of chemotherapy administration every 4 weeks, for a total of 5 FMD cycles during neoadjuvant chemotherapy (4 times dose dense Adriamycin plus cyclophosphamide followed by 12 times weekly paclitaxel). Patients will be supported and monitored by a centralized dietician during each FMD cycle. An additional MRI will be conducted after 4ddAC to evaluate efficacy mid-way through chemotherapy. Furthermore, an optional tumor biopsy will be done to determine the immunomodulatory effects of the FMD which will be determined using cutting-edge RNA sequencing and multispectral imaging techniques. Pathological response will be graded on the surgical specimen according to Miller & Payne 4/5, indicating a 90-100% tumor cell loss. Radiological response will be determined by MRI (RECIST1.1). In total 240 patients are needed to detect an increase from 20% to 37.5% in pathological response rate in the FMD arm allowing for 15% dropout, using a two-sided Z test with pooled variance and 5% significance level with a power of 80%. Randomization will be performed in Castor EDC with variable block sizes and stratified by clinical stage (stadium II versus III) and BMI (< 25kg/m2 versus ≥ 25kg/m2). Inclusion criteria are HR+, HER2- breast cancer stage II-III, BMI > 18.5 kg/m2, WHO status 0-2, without a history of malignancy in the past 5 years, invasive or ipsilateral non-invasive breast cancer. Patients will be excluded in case of pregnancy, lactation or metabolic disease that affects gluconeogenesis and adaptability to FMD. The trial is supported by grants from the Dutch Cancer Society and WCRF. The study is open since February 2023 for inclusion for 4 years and plans to recruit 240 patients in total in 20 participating hospitals in the Netherlands. Clinical trial information: NCT05503108 .

Find: Immunomodulation after a fasting mimicking diet analyzed by Nanostring and multispectral flowcytometry in healthy volunteers: A pilot study.

e14560 Background: An increasing number of clinical trials, including our randomized phase III trial (NCT05503108), are investigating the potential benefit of short-term fasting using a fasting mimicking diet (FMD) on the anti-tumor effect of chemotherapy. Preclinical research suggests that FMD promotes infiltration of CD8+ T-cells, while decreasing regulatory T-cells (Treg). Extensive immunoprofiling is needed to investigate the immunomodulatory effects of FMD alone and guide ongoing translational trials on FMD as an adjunct to chemotherapy in patients with cancer. Methods: FIND is a prospective pilot study, which included 9 healthy volunteers who followed a 4-day FMD every 3 weeks, 2 times (NCT04833439). Peripheral blood samples were collected in a fed state (baseline), after overnight fasting (overnight) and directly after the second cycle of FMD (final). Metabolic markers (IGF1, glucose, ketone bodies) to ensure ketosis were measured in the plasma. The RNA from white blood cells was isolated and transcriptomics was performed with the Nanostring IO360 human Pancancer panel, and analyzed by nSolver Advanced analysis, Graphpad Prism and String analysis. Statistical testing was done using Wilcoxon signed-rank tests in R-studio. In addition, 40-plex spectral AURORA flow cytometry for immune cell composition with FlowJo analysis was performed. This study was approved by the local Medical ethical committee (NL76033.058.21). Results: Paired analysis of the Nanostring data identified 56 differentially expressed genes (DEG) comparing baseline and final samples initially, of which 19 genes showed at least 1.2 log2 fold-change in at least half of the participants. String analysis indicated that the DEGs were associated with fatty-acid metabolism, T-cell and myeloid cell function and immune regulatory pathways. Cell typing showed an increase in cytotoxic NK CD56dim cells (p=0.031) and a decrease in Tregs decreased (p=0.063) after FMD. In-depth analysis of gene specific pathway enrichment and immune cell profiling by flow cytometry will be presented. Conclusions: The expression profile of 56 genes in circulating immune cells changed after 2 FMD cycles, next to fatty-acid metabolism affecting the composition and function of both T-cells and myeloid cells and their regulation. Altogether, FMD induced immunological changes that could support cancer treatment and guide immunomonitoring in future trials evaluating the effects of FMD in cancer treatment. Clinical trial information: NCT04833439 .

Six-month periodic fasting does not affect somatosensory nerve function in type 2 diabetes patients.

Current strategies for preventing diabetic sensorimotor polyneuropathy (DSPN) are limited mainly to glucose control but rapid decrease of glycemia can lead to acute onset or worsening of DSPN. The aim of this study was to examine the effects of periodic fasting on somatosensory nerve function in patients with type 2 diabetes (T2D). Somatosensory nerve function was assessed in thirty-one patients with T2D (HbA1c 7.8 ± 1.3% [61.4 ± 14.3 mmol/mol]) before and after a six-month fasting-mimicking diet (FMD; n=14) or a control Mediterranean diet (M-diet; n=17). Neuropathy disability score (NDS), neuropathy symptoms score (NSS), nerve conduction velocity and quantitative sensory testing (QST) were analyzed. 6 participants of the M-Diet group and 7 of the FMD group underwent diffusion-weighted high-resolution magnetic resonance neurography (MRN) of the right leg before and after the diet intervention. Clinical neuropathy scores did not differ between study groups at baseline (64% in the M-Diet group and 47% in the FMD group had DSPN) and no change was found after intervention. The differences in sensory NCV and sensory nerve action potential (SNAP) of sural nerve were comparable between study groups. Motor NCV of tibial nerve decreased by 12% in the M-Diet group (P=0.04), but did not change in the FMD group (P=0.39). Compound motor action potential (CMAP) of tibial nerve did not change in M-Diet group (P=0.8) and increased in the FMD group by 18% (P=0.02). Motor NCV and CMAP of peroneal nerve remained unchanged in both groups. In QST M-diet-group showed a decrease by 45% in heat pain threshold (P=0.02), FMD group showed no change (P=0.50). Changes in thermal detection, mechanical detection and mechanical pain did not differ between groups. MRN analysis showed stable fascicular nerve lesions irrespective of the degree of structural pathology. Fractional anisotropy and T2-time did not change in both study groups, while a correlation with the clinical degree of DSPN could be confirmed for both. Our study shows that six-month periodic fasting was safe in preserving nerve function and had no detrimental effects on somatosensory nerve function in T2D patients. https://drks.de/search/en/trial/DRKS00014287, identifier DRKS00014287.

Persistent Degradation of HER2 Protein by Hybrid nanoPROTAC for Programmed Cell Death.

Proteolysis-targeting chimera (PROTAC) has emerged as a promising strategy for degrading proteins of interest. Peptide-based PROTACs offer several advantages over small-molecule-based PROTACs, such as high specificity, low toxicity, and large protein-protein interaction surfaces. However, peptide-based PROTACs have several intrinsic shortcomings that strongly limit their application including poor cell permeability and low stability and potency. Herein, we designed a nanosized hybrid PROTAC (GNCTACs) to target and degrade human epidermal growth factor receptor 2 (HER2) in tumor cells. Gold nanoclusters (GNCs) were utilized to connect HER2-targeting peptides and cereblon (CRBN)-targeting ligands. GNCTACs could overcome the intrinsic barriers of peptide-based PROTACs, efficiently delivering HER2-targeting peptides in the cytoplasm and protecting them from degradation. Furthermore, a fasting-mimicking diet was applied to enhance the cellular uptake and proteasome activity. Consequently, more than 95% of HER2 in SKBR3 cells was degraded by GNCTACs, and the degradation lasted for at least 72 h, showing a catalytic-like reaction.

Fasting-mimicking diet synergizes with ferroptosis against quiescent, chemotherapy-resistant cells.

More than ten randomized clinical trials are being tested to evaluate the efficacy, effectiveness and safety of a fasting-mimicking diet (FMD) combined with different antitumor agents. UMI-mRNA sequencing, Cell-cycle analysis, Label retention, metabolomics, Multilabeling etal. were used to explore mechanisms. A tandem mRFP-GFP-tagged LC3B, Annexin-V-FITC Apoptosis, TUNEL, H&E, Ki-67 and animal model was used to search for synergistic drugs. Here we showed that fasting or FMD retards tumor growth more effectively but does not increase 5-fluorouracil/oxaliplatin (5-FU/OXA) sensitivity to apoptosis invitro and invivo. Mechanistically, we demonstrated that CRC cells would switch from an active proliferative to a slow-cycling state during fasting. Furthermore, metabolomics shows cell proliferation was decreased to survive nutrient stress invivo, as evidenced by a low level of adenosine and deoxyadenosine monophosphate. CRC cells would decrease proliferation to achieve increased survival and relapse after chemotherapy. In addition, these fasting-induced quiescent cells were more prone to develop drug-tolerant persister (DTP) tumor cells postulated to be responsible for cancer relapse and metastasis. Then, UMI-mRNA sequencing uncovered the ferroptosis pathway as the pathway most influenced by fasting. Combining fasting with ferroptosis inducer treatment leads to tumor inhibition and eradication of quiescent cells by boosting autophagy. Our results suggest that ferroptosis could improve the antitumor activity of FMD+chemotherapy and highlight a potential therapeutic opportunity to avoid DTP cells-driven tumor relapse and therapy failure. A full list of funding bodies can be found in the Acknowledgements section.

82P Cyclic fasting-mimicking diet as a strategy to improve the efficacy of standard antitumor therapies in cancer patients

82P Cyclic fasting-mimicking diet as a strategy to improve the efficacy of standard antitumor therapies in cancer patients