mTORC1 and BMP-Smad1/5 regulation of serum-stimulated myotube hypertrophy: a role for autophagy.

Protein synthesis regulation is critical for skeletal muscle hypertrophy, yet other established cellular processes are necessary for growth-related cellular remodeling. Autophagy has a well-acknowledged role in muscle quality control, but evidence for its role in myofiber hypertrophy remains equivocal. Both mammalian target of rapamycin complex I (mTORC1) and bone morphogenetic protein (BMP)-Smad1/5 (Sma and Mad proteins from Caenorhabditis elegans and Drosophila, respectively) signaling are reported regulators of myofiber hypertrophy; however, gaps remain in our understanding of how this regulation is integrated with growth processes and autophagy regulation. Therefore, we investigated the mTORC1 and Smad1/5 regulation of protein synthesis and autophagy flux during serum-stimulated myotube growth. Chronic serum stimulation experiments were performed on day 5 differentiated C2C12 myotubes incubated in differentiation medium [2% horse serum (HS)] or growth medium [5% fetal bovine serum (FBS)] for 48 h. Rapamycin or LDN193189 was dosed for 48 h to inhibit mTORC1 and BMP-Smad1/5 signaling, respectively. Acute serum stimulation was examined in day 7 differentiated myotubes. Protein synthesis was measured by puromycin incorporation. Bafilomycin A1 and immunoblotting for LC3B were used to assess autophagy flux. Chronic serum stimulation increased myotube diameter 22%, total protein 21%, total RNA 100%, and Smad1/5 phosphorylation 404% and suppressed autophagy flux. Rapamycin, but not LDN193189, blocked serum-induced myotube hypertrophy and the increase in total RNA. Acute serum stimulation increased protein synthesis 111%, Smad1/5 phosphorylation 559%, and rpS6 phosphorylation 117% and suppressed autophagy flux. Rapamycin increased autophagy flux during acute serum stimulation. These results provide evidence for mTORC1, but not BMP-Smad1/5, signaling being required for serum-induced myotube hypertrophy and autophagy flux by measuring LC3BII/I expression. Further investigation is warranted to examine the role of autophagy flux in myotube hypertrophy.NEW & NOTEWORTHY The present study demonstrates that myotube hypertrophy caused by chronic serum stimulation requires mammalian target of rapamycin complex 1 (mTORC1) signaling but not bone morphogenetic protein (BMP)-Smad1/5 signaling. The suppression of autophagy flux was associated with serum-induced myotube hypertrophy and mTORC1 regulation of autophagy flux by measuring LC3BII/I expression. Rapamycin is widely investigated for beneficial effects in aging skeletal muscle and sarcopenia; our results provide evidence that rapamycin can regulate autophagy-related signaling during myotube growth, which could benefit skeletal muscle functional and metabolic health.

Recovery from FOLFOX chemotherapy-induced systemic and skeletal muscle metabolic dysfunction in mice.

FOLFOX (5-fluorouracil, leucovorin, oxaliplatin) chemotherapy is used to treat colorectal cancer and can acutely induce metabolic dysfunction. However, the lasting effects on systemic and skeletal muscle metabolism after treatment cessation are poorly understood. Therefore, we investigated the acute and lasting effects of FOLFOX chemotherapy on systemic and skeletal muscle metabolism in mice. Direct effects of FOLFOX in cultured myotubes were also investigated. Male C57BL/6J mice completed four cycles (acute) of FOLFOX or PBS. Subsets were allowed to recover for 4 wk or 10 wk. Comprehensive Laboratory Animal Monitoring System (CLAMS) metabolic measurements were performed for 5 days before study endpoint. C2C12 myotubes were treated with FOLFOX for 24 hr. Acute FOLFOX attenuated body mass and body fat accretion independent of food intake or cage activity. Acute FOLFOX decreased blood glucose, oxygen consumption (V̇o2), carbon dioxide production (V̇co2), energy expenditure, and carbohydrate (CHO) oxidation. Deficits in V̇o2 and energy expenditure remained at 10 wk. CHO oxidation remained disrupted at 4 wk but returned to control levels after 10 wk. Acute FOLFOX reduced muscle COXIV enzyme activity, AMPK(T172), ULK1(S555), and LC3BII protein expression. Muscle LC3BII/I ratio was associated with altered CHO oxidation (r = 0.75, P = 0.03). In vitro, FOLFOX suppressed myotube AMPK(T172), ULK1(S555), and autophagy flux. Recovery for 4 wk normalized skeletal muscle AMPK and ULK1 phosphorylation. Our results provide evidence that FOLFOX disrupts systemic metabolism, which is not readily recoverable after treatment cessation. FOLFOX effects on skeletal muscle metabolic signaling did recover. Further investigations are warranted to prevent and treat FOLFOX-induced metabolic toxicities that negatively impact survival and life quality of patients with cancer.NEW & NOTEWORTHY The present study demonstrates that FOLFOX chemotherapy induces long-lasting deficits in systemic metabolism. Interestingly, FOLFOX modestly suppressed skeletal muscle AMPK and autophagy signaling in vivo and in vitro. The FOLFOX-induced suppression of muscle metabolic signaling recovered after treatment cessation, independent of systemic metabolic dysfunction. Future research should investigate if activating AMPK during treatment can prevent long-term toxicities to improve health and quality of life of patients with cancer and survivors.

Short duration treadmill exercise improves physical function and skeletal muscle mitochondria protein expression after recovery from FOLFOX chemotherapy in male mice.

FOLFOX (5-FU, leucovorin, oxaliplatin) is a chemotherapy treatment for colorectal cancer which induces toxic side effects involving fatigue, weakness, and skeletal muscle dysfunction. There is a limited understanding of the recovery from these toxicities after treatment cessation. Exercise training can improve chemotherapy-related toxicities. However, how exercise accelerates recovery and the dose required for these benefits are not well examined. The purpose of this study was to examine the effect of exercise duration on physical function, muscle mass, and mitochondria protein expression during the recovery from FOLFOX chemotherapy. 12-week-old male mice were administered four cycles of either PBS or FOLFOX over 8-weeks. Outcomes were assessed after the fourth cycle and after either 4 (short-term; STR) or 10 weeks (long-term; LTR) recovery. Subsets of mice performed 14 sessions (6 d/wk, 18 m/min, 5% grade) of 60 min/d (long) or 15 min/d (short duration) treadmill exercise during STR. Red and white gastrocnemius mRNA and protein expression were examined. FOLFOX treatment decreased run time (RT) (-53%) and grip strength (GS) (-9%) compared to PBS. FOLFOX also reduced muscle OXPHOS complexes, COXIV, and VDAC protein expression. At LTR, FOLFOX RT (-36%) and GS (-16%) remained reduced. Long- and short-duration treadmill exercise improved RT (+58% and +56%) without restoring GS in FOLFOX mice. Both exercise durations increased muscle VDAC and COXIV expression in FOLFOX mice. These data provide evidence that FOLFOX chemotherapy induces persistent deficits in physical function that can be partially reversed by short-duration aerobic exercise.

Validity of the Handheld Doppler to Determine Lower-Limb Blood Flow Restriction Pressure for Exercise Protocols.

Laurentino, GC, Loenneke, JP, Mouser, JG, Buckner, SL, Counts, BR, Dankel, SJ, Jessee, MB, Mattocks, KT, Iared, W, Tavares, LD, Teixeira, EL, and Tricoli, V. Validity of the handheld Doppler to determine lower-limb blood flow restriction pressure for exercise protocols. J Strength Cond Res 34(9): 2693-2696, 2020-Handheld (HH) Doppler is frequently used for determining the arterial occlusion pressure during blood flow restriction exercises; however, it is unknown whether the blood flow is occluded when the auscultatory signal is no longer present. The purpose of this study was to assess the validity between the HH Doppler and the Doppler ultrasound (US) measurements for determining the arterial occlusion pressure in healthy men. Thirty-five participants underwent 2 arterial occlusion pressure measurements. In the first measure, a pressure cuff (17.5 cm wide) was placed at the most proximal region of the thigh and the pulse of posterior tibial artery was detected using an HH Doppler probe. The cuff was inflated until the auscultatory pulse was no longer detected. After 10 minutes of rest, the procedure was repeated with the Doppler US probe placed on the superficial femoral artery. The cuff was inflated up to the point at which the femoral arterial blood flow was interrupted. The point at which the auscultatory pulse and blood flow were no longer detected was deemed the arterial occlusion pressure. There were no significant differences in arterial occlusion pressure level between the HH Doppler and the Doppler US (133 [±18] vs. 135 [±17] mm Hg, p = 0.168). There was a significant correlation (r = 0.938, p = 0.168), reasonable agreement, and a total error of the estimate of 6.0 mm Hg between measurements. Arterial occlusion pressure level determined by the HH Doppler and the Doppler US was similar, providing evidence that the HH Doppler is a valid and practical method.

The regulation of skeletal muscle fatigability and mitochondrial function by chronically elevated interleukin-6.

NEW FINDINGS: What is the central question of this study? Interleukin-6 has been associated with muscle mass and metabolism in both physiological and pathological conditions. A causal role for interleukin-6 in the induction of fatigue and disruption of mitochondrial function has not been determined. What is the main finding and its importance? We demonstrate that chronically elevated interleukin-6 increased skeletal muscle fatigability and disrupted mitochondrial content and function independent of changes in fibre type and mass. ABSTRACT: Interleukin-6 (IL-6) can initiate intracellular signalling in skeletal muscle by binding to the IL-6-receptor and interacting with the transmembrane gp130 protein. Circulating IL-6 has established effects on skeletal muscle mass and metabolism in both physiological and pathological conditions. However, the effects of circulating IL-6 on skeletal muscle function are not well understood. The purpose of this study was to determine whether chronically elevated systemic IL-6 was sufficient to disrupt skeletal muscle force, fatigue and mitochondrial function. Additionally, we examined the role of muscle gp130 signalling during overexpression of IL-6. Systemic IL-6 overexpression for 2 weeks was achieved by electroporation of an IL-6 overexpression plasmid or empty vector into the quadriceps of either C57BL/6 (WT) or skeletal muscle gp130 knockout (KO) male mice. Tibialis anterior muscle in situ functional properties and mitochondrial respiration were determined. Interleukin-6 accelerated in situ skeletal muscle fatigue in the WT, with a 18.5% reduction in force within 90 s of repeated submaximal contractions and a 7% reduction in maximal tetanic force after 5 min. There was no difference between fatigue in the KO and KO+IL-6. Interleukin-6 reduced WT muscle mitochondrial respiratory control ratio by 36% and cytochrome c oxidase activity by 42%. Interleukin-6 had no effect on either KO respiratory control ratio or cytochrome c oxidase activity. Interleukin-6 also had no effect on body weight, muscle mass or tetanic force in either genotype. These results provide evidence that 2 weeks of elevated systemic IL-6 is sufficient to increase skeletal muscle fatigability and decrease muscle mitochondrial content and function, and these effects require muscle gp130 signalling.

Muscle adaptations following 21 consecutive days of strength test familiarization compared with traditional training.

INTRODUCTION: Large increases in 1-repetition maximum (1RM) strength have been demonstrated from repeated testing, but it is unknown whether these increases can be augmented by resistance training. METHODS: Five trained individuals performed a 1RM test and maximal voluntary isometric contraction (MVC) for unilateral elbow flexion exercise on 1 arm (testing arm), while the other arm performed a 1RM test and MVC, in addition to 3 sets of exercise (70% 1RM) (training arm), for 21 straight days. RESULTS: Although only the training arm had increased muscle thickness [mean 0.28 cm, 95% confidence interval (CI) 0.22-0.33 cm], 1RM strength increased similarly in the training (2.2 kg, 95% CI 0.9-3.4 kg; P = 0.008) and testing (1.9 kg, 95% CI 0.5-3.2 kg; P = 0.019) arms. CONCLUSION: Increases in 1RM strength from resistance training are related to the specificity of exercise and are likely driven by mechanisms other than muscle growth. Muscle Nerve 56: 307-314, 2017.

Muscle growth: To infinity and beyond?

Strength increases following training are thought to be influenced first by neural adaptions and second by large contributions from muscle growth. This is based largely on the idea that muscle growth is a slow process and that a plateau in muscle growth would substantially hinder long-term increases in strength. This Review examines the literature to determine the time course of skeletal muscle growth in the upper and lower body and to determine whether and when muscle growth plateaus. Studies were included if they had at least 3 muscle size time points, involved participants 18 years or older, and used a resistance training protocol. Muscle growth occurs sooner than had once been hypothesized, and this adaptation is specific to the muscle group. Furthermore, the available studies indicate that the muscle growth response will plateau, and additional growth is not likely to occur appreciably beyond this initial plateau. However, the current study durations are a limitation. Muscle Nerve 56: 1022-1030, 2017.

A tale of three cuffs: the hemodynamics of blood flow restriction.

INTRODUCTION: The blood flow response to relative levels of blood flow restriction (BFR) across varying cuff widths is not well documented. With the variety of cuff widths and pressures reported in the literature, the effects of different cuffs and pressures on blood flow require investigation. PURPOSE: To measure blood pressure using three commonly used BFR cuffs, examine possible venous/arterial restriction pressures, and measure hemodynamic responses to relative levels of BFR using these same cuffs. METHODS: 43 participants (Experiment 1, brachial artery blood pressure assessed) and 38 participants (Experiment 2, brachial artery blood flow assessed using ultrasound, cuff placed at proximal portion of arm) volunteered for this study. RESULTS: Blood pressure measurement was higher in the 5 cm cuff than in the 10 and 12 cm cuffs. Sub-diastolic relative pressures appear to occur predominantly at <60% of arterial occlusion pressure (AOP). Blood flow under relative levels of restriction decreases in a non-linear fashion, with minimal differences between cuffs [resting: 50.3 (44.2) ml min-1; 10% AOP: 42.0 (36.8); 20%: 33.6 (28.6); 30%: 23.6 (20.4); 40%: 17.1 (15.9); 50%: 12.5 (9.4); 60%: 11.5 (8.1); 70%: 11.4 (7.0); 80%: 10.3 (6.3); 90%: 7.9 (4.8); 100%: 1.5 (2.9)]. Peak blood velocity remains relatively constant until higher levels (>70% of AOP) are surpassed. Calculated mean shear rate decreases in a similar fashion as blood flow. CONCLUSIONS: Under relative levels of restriction, pressures from 40 to 90% of AOP appear to decrease blood flow to a similar degree in these three cuffs. Relative pressures appear to elicit a similar blood flow stimulus when accounting for cuff width and participant characteristics.

Differentiating swelling and hypertrophy through indirect assessment of muscle damage in untrained men following repeated bouts of resistance exercise.

PURPOSE: To examine the swelling response and other markers of muscle damage throughout the early portions of a training program (Experiment 1). We also determined if a "swollen" muscle could swell further following additional exercise (Experiment 2). METHODS: Nine males performed four sets of biceps curls (or time-matched rest on control arm) at 70% of their one-repetition maximum three times over 8 days. Muscle thickness and torque were measured before and after exercise as well as on the days in between. Soreness was measured at the beginning of each day (Experiment 1). On the final day (Experiment 2), participants performed two bouts of exercise, followed by additional measures of muscle thickness. RESULTS: Following three bouts of exercise, muscle thickness was elevated over baseline (mean of visit 9 pre to visit 2 pre, 95% CI) at the 50% [0.21 (0.07, 0.34) cm], 60% [0.21 (0.02, 0.39) cm], and 70% [0.21 (0.06, 0.36) cm] sites. However, differences from a non-exercise control were only observed immediately following bouts of exercise (indicative of acute swelling). Torque was lower at every time point following the first bout of exercise and remained suppressed relative to pre at visit 9 [-6.1 (-11.7, -0.47 Nm] in the experimental arm. Experiment 2 found that a swollen muscle could not appreciably swell more. CONCLUSION: Resting levels of muscle thickness do not appear to change beyond what occurs following the first naïve bout of exercise. Also, the acute swelling response may be used to differentiate swelling from muscle growth.

Objectively-Measured Free-Living Physical Activity and Heart Rate Recovery.

The purpose of this study was to examine the association of free-living, objectively-measured physical activity on treadmill-based heart rate recovery (HRR), a parameter known to associate with morbidity and mortality. Data was used from 2003 to 2004 NHANES. Physical activity was assessed via accelerometry, with HRR recovery assessed from a treadmill-based test. Heart rate recovery minute 1 (HRR1) and minute 2 (HRR2) were calculated. After adjustment, light and vigorous-intensity free-living physical activity, respectively, were associated with HRR1 (ßadjusted = 0.69, 95% CI 0.22-1.14; ßadjusted 1.94, 95% CI 0.01-3.9) and HRR2 (ßadjusted = 0.99, 95% CI 0.35-1.62; ßadjusted = 5.88, 95% CI 2.63-9.12). Moderate physical activity was not associated with HRR1 (ßadjusted = 0.60, 95% CI -0.41 to 1.62), but was with HRR2 (ßadjusted = 2.28, 95% CI 1.27-3.28). As free-living physical activity intensity increased, there was a greater association with HRR. This finding may provide mechanistic insight of previous research observations demonstrating intensity-specific effects of physical activity on various health outcomes.

The problem Of muscle hypertrophy: Revisited.

In this paper we revisit a topic originally discussed in 1955, namely the lack of direct evidence that muscle hypertrophy from exercise plays an important role in increasing strength. To this day, long-term adaptations in strength are thought to be primarily contingent on changes in muscle size. Given this assumption, there has been considerable attention placed on programs designed to allow for maximization of both muscle size and strength. However, the conclusion that a change in muscle size affects a change in strength is surprisingly based on little evidence. We suggest that these changes may be completely separate phenomena based on: (1) the weak correlation between the change in muscle size and the change in muscle strength after training; (2) the loss of muscle mass with detraining, yet a maintenance of muscle strength; and (3) the similar muscle growth between low-load and high-load resistance training, yet divergent results in strength. Muscle Nerve 54: 1012-1014, 2016.

Influence of relative blood flow restriction pressure on muscle activation and muscle adaptation.

INTRODUCTION: The aim of this study was to investigate the acute and chronic skeletal muscle response to differing levels of blood flow restriction (BFR) pressure. METHODS: Fourteen participants completed elbow flexion exercise with pressures from 40% to 90% of arterial occlusion. Pre/post torque measurements and electromyographic (EMG) amplitude of each set were quantified for each condition. This was followed by a separate 8-week training study of the effect of high (90% arterial occlusion) and low (40% arterial occlusion) pressure on muscle size and function. RESULTS: For the acute study, decreases in torque were similar between pressures [-15.5 (5.9) Nm, P = 0.344]. For amplitude of the first 3 and last 3 reps there was a time effect. After training, increases in muscle size (10%), peak isotonic strength (18%), peak isokinetic torque (180°/s = 23%, 60°/s = 11%), and muscular endurance (62%) changed similarly between pressures. CONCLUSION: We suggest that higher relative pressures may not be necessary when exercising under BFR.

Post-exercise blood flow restriction attenuates muscle hypertrophy.

PURPOSE: Applying blood flow restriction during low-load resistance training has been shown to augment muscle hypertrophy which has been attributed to metabolic accumulation. It remains unknown, however, whether metabolites can augment muscle growth when maintained post-exercise. METHODS: Thirteen untrained individuals (6 males and 7 females) performed 24 training sessions. The control arm performed one set of elbow flexion (70 % 1RM) to volitional fatigue, while the experimental arm performed the same protocol immediately followed by 3 min of blood flow restriction (70 % arterial occlusion). Muscle growth (ultrasound) was measured at 50, 60, and 70 % of the distance between the lateral epicondyle and acromion process. RESULTS: Both conditions completed the same exercise volume [3678 (95 % CI 2962, 4393) vs. 3638 kg (95 % CI 2854, 4423)]. There was a condition by time interaction (p = 0.031) demonstrating an attenuation of muscle growth at the 60 % site in the experimental [pre 3.1 (95 % CI 2.8, 3.5), post 3.1 (95 % CI 2.7, 3.5) cm] vs. control [pre 3.1 (95 % CI 2.6, 3.6), post 3.3 (95 % CI 2.8, 3.7) cm] condition. Muscle growth at the 50 % and 70 % sites was similar at the group level, although there were attenuations at the individual level. Exploratory analyses of pre-post mean (95 % CI) changes in muscle thickness suggested that this attenuation in the experimental condition occurred only in females [50 % site 0.0 (-0.2, 0.0) cm; 60 % site -0.1 (-0.3, 0.0) cm; 70 % site 0.0 (-0.1, 0.1) cm]. CONCLUSIONS: The application of blood flow restriction post high-load training did not augment muscle growth for either sex, and appeared to attenuate muscle growth among females.

Mouse skeletal muscle adaptations to different durations of treadmill exercise after the cessation of FOLFOX chemotherapy.

FOLFOX (5-fluorouracil, leucovorin, oxaliplatin) chemotherapy is a treatment for colorectal cancer that can induce persistent fatigue and metabolic dysfunction. Regular exercise after chemotherapy cessation is widely recommended for cancer patients and has been shown to improve fatigue resistance in mice. However, gaps remain in understanding whether the early systemic and skeletal muscle adaptations to regular exercise are altered by prior FOLFOX chemotherapy treatment. Furthermore, the effects of exercise duration on early metabolic and skeletal muscle transcriptional adaptations are not fully established. Purpose: Investigate the effects of prior FOLFOX chemotherapy treatment on the early adaptations to repeated short- or long-duration treadmill exercise, including the fasting regulation of circulating metabolic regulators, skeletal muscle COXIV activity and myokine/exerkine gene expression in male mice. Methods: Male C57BL6/J mice completed 4 cycles of FOLFOX or PBS and were allowed to recover for 4-weeks. Subsets of mice performed 14 sessions (6 d/wk, 18 m/min, 5% grade) of short- (10 min/d) or long-duration (55 min/d) treadmill exercise. Blood plasma and muscle tissues were collected 48-72 h after the last exercise bout for biochemical analyses. Results: Long-duration exercise increased fasting plasma osteocalcin, LIF, and IL-6 in healthy PBS mice, and these changes were ablated by prior FOLFOX treatment. Slow-oxidative soleus muscle COXIV activity increased in response to long-duration exercise in PBS mice, which was blocked by prior FOLFOX treatment. Fast-glycolytic plantaris muscle COXIV activity increased with short-duration exercise independent of FOLFOX administration. There was a main effect for long-duration exercise to increase fasting muscle IL-6 and COXIV mRNA expression independent of FOLFOX. FOLFOX administration reduced muscle IL-6, LIF, and BDNF mRNA expression irrespective of long-duration exercise. Interestingly, short-duration exercise suppressed the FOLXOX induction of muscle myostatin mRNA expression. Conclusion: FOLFOX attenuated early exercise adaptations related to fasting circulating osteocalcin, LIF, and IL-6. However, prior FOLFOX treatment did not alter the exercise adaptations of plantaris muscle COXIV activity and plasma adiponectin. An improved understanding of mechanisms underlying exercise adaptations after chemotherapy will provide the basis for successfully treating fatigue and metabolic dysfunction in cancer survivors.

Early-Onset Physical Inactivity and Metabolic Dysfunction in Tumor-bearing Mice Is Associated with Accelerated Cachexia.

METHODS: Male C57BL/6J mice (12 wk of age) were injected with 1 × 106 LLC cells or phosphate-buffered saline (PBS) subcutaneously in the right flank, and tissue was collected 26-28 d after cell injection. Tumor volume was measured every 5 d throughout the study to calculate the tumor growth rate. Fifteen days after tumor inoculation, a subset of PBS (n = 11) and LLC (n = 16) mice were individually housed in metabolic Comprehensive Laboratory Animal Monitoring System cages for 5 d. RESULTS: LLC mice exhibited greater body weight loss (-5.1%), decreased muscle mass (-7%), decreased fat mass (-22%), and increased plasma interleukin-6 (212%) compared with PBS mice. Before the onset of cachexia, total cage activity was decreased in tumor-bearing mice. Cage activity was negatively associated with tumor mass and positively associated with hindlimb muscle mass. In addition, LLC mice had greater lipid oxidation than PBS mice. CONCLUSIONS: LLC mice exhibit early-onset physical inactivity and altered systemic lipid oxidation, which are associated with the eventual development of cachexia.

PKC agonism restricts innate immune suppression, promotes antigen cross-presentation and synergizes with agonistic CD40 antibody therapy to activate CD8+ T cells in breast cancer.

Myeloid-derived suppressor cells (MDSCs) are an immature innate cell population that expands in pathological conditions such as cancer and suppresses T cells via production of immunosuppressive factors. Conversely, efficient cytotoxic T cell priming is dependent on the ability of antigen-presenting cells (APCs) to cross-present tumor antigens to CD8+ T cells, a process that requires a specific subtype of dendritic cells (DCs) called conventional DC1 (cDC1) which are often dysfunctional in cancer. One way to activate cDC1 is ligation of CD40 which is abundantly expressed by myeloid cells and its agonism leads to myeloid cell activation. Thus, targeting MDSCs while simultaneously expanding cross-presenting DCs represents a promising strategy that, when combined with agonistic CD40, may result in long-lasting protective immunity. In this study, we investigated the effect of PKC agonists PEP005 and prostratin on MDSC expansion, differentiation, and recruitment to the tumor microenvironment. Our findings demonstrate that PKC agonists decreased MDSC expansion from hematopoietic progenitors and induced M-MDSC differentiation to an APC-like phenotype that expresses cDC1-related markers via activation of the p38 mitogen-activated protein kinase (MAPK) pathway. Simultaneously, PKC agonists favored cDC1 expansion at the expense of cDC2 and plasmacytoid DCs (pDC). Functionally, PKC agonists blunted MDSC suppressive activity and enhanced MDSC cross-priming capacity both in vitro and in vivo. Finally, combination of PKC agonism with agonistic CD40 mAb resulted in a marked reduction in tumor growth with a significant increase in intratumoral activated CD8+ T cells and tissue-resident memory CD8+ T cells in a syngeneic breast cancer mouse model. In sum, this work proposes a novel promising strategy to simultaneously target MDSCs and promote APC function that may have highly impactful clinical relevance in cancer patients.

Response to immune checkpoint blockade improved in pre-clinical model of breast cancer after bariatric surgery.

Bariatric surgery is a sustainable weight loss approach, including vertical sleeve gastrectomy (VSG). Obesity exacerbates tumor growth, while diet-induced weight loss impairs progression. It remains unknown how bariatric surgery-induced weight loss impacts cancer progression or alters response to therapy. Using a pre-clinical model of obesity followed by VSG or diet-induced weight loss, breast cancer progression and immune checkpoint blockade therapy were investigated. Weight loss by VSG or weight-matched dietary intervention before tumor engraftment protected against obesity-exacerbated tumor progression. However, VSG was not as effective as diet in reducing tumor burden despite achieving similar weight and adiposity loss. Leptin did not associate with changes in tumor burden; however, circulating IL-6 was elevated in VSG mice. Uniquely, VSG tumors displayed elevated inflammation and immune checkpoint ligand PD-L1+ myeloid and non-immune cells. VSG tumors also had reduced T lymphocytes and markers of cytolysis, suggesting an ineffective anti-tumor microenvironment which prompted investigation of immune checkpoint blockade. While obese mice were resistant to immune checkpoint blockade, anti-PD-L1 potently impaired tumor progression after VSG through improved anti-tumor immunity. Thus, in formerly obese mice, surgical weight loss followed by immunotherapy reduced breast cancer burden. Finally, we compared transcriptomic changes in adipose tissue after bariatric surgery from patients and mouse models. A conserved bariatric surgery-associated weight loss signature (BSAS) was identified which significantly associated with decreased tumor volume. Findings demonstrate conserved impacts of obesity and bariatric surgery-induced weight loss pathways associated with breast cancer progression.

As the number of people classified as obese rises globally, so do obesity-related health risks. Studies show that people diagnosed with obesity have inflammation that contributes to tumor growth and their immune system is worse at detecting cancer cells. But weight loss is not currently used as a strategy for preventing or treating cancer. Surgical procedures for weight loss, also known as 'bariatric surgeries', are becoming increasingly popular. Recent studies have shown that individuals who lose weight after these treatments have a reduced risk of developing tumors. But how bariatric surgery directly impacts cancer progression has not been well studied: does it slow tumor growth or boost the anti-tumor immune response? To answer these questions, Sipe et al. compared breast tumor growth in groups of laboratory mice that were obese due to being fed a high fat diet. The first group of mice lost weight after undergoing a bariatric surgery in which part of their stomach was removed. The second lost the same amount of weight but after receiving a restricted diet, and the third underwent a fake surgery and did not lose any weight. The experiments found that surgical weight loss cuts breast cancer tumor growth in half compared with obese mice. But mice who lost the same amount of weight through dietary restrictions had even less tumor growth than surgically treated mice. The surgically treated mice who lost weight had more inflammation than mice in the two other groups, and had increased amounts of proteins and cells that block the immune response to tumors. Giving the surgically treated mice a drug that enhances the immune system's ability to detect and destroy cancer cells reduced inflammation and helped shrink the mice's tumors. Finally, Sipe et al. identified 54 genes which were turned on or off after bariatric surgery in both mice and humans, 11 of which were linked with tumor size. These findings provide crucial new information about how bariatric surgery can impact cancer progression. Future studies could potentially use the conserved genes identified by Sipe et al. to develop new ways to stimulate the anti-cancer benefits of weight loss without surgery.

Wheel running improves fasting-induced AMPK signaling in skeletal muscle from tumor-bearing mice.

Disruptions to muscle protein turnover and metabolic regulation contribute to muscle wasting during the progression of cancer cachexia. The initiation of cachexia is also associated with decreased physical activity. While chronic muscle AMPK activation occurs during cachexia progression in ApcMin/+ (MIN) mice, a preclinical cachexia model, the understanding of muscle AMPK's role during cachexia initiation is incomplete. Therefore, we examined if voluntary wheel exercise could improve skeletal muscle AMPK signaling in pre-cachectic MIN mice. Next, we examined muscle AMPK's role in aberrant catabolic signaling in response to a 12-h fast in mice initiating cachexia. Male C57BL/6 (B6: N = 26) and MIN (N = 29) mice were subjected to ad libitum feeding, 12-h fast, or 4 wks. of wheel access and then a 12-h fast during the initiation of cachexia. Male tamoxifen-inducible skeletal muscle AMPKα1 α2 (KO) knockout mice crossed with ApcMin/+ and floxed controls were examined (WT: N = 8, KO: N = 8, MIN: N = 10, MIN KO: N = 6). Male mice underwent a 12-h fast and the gastrocnemius muscle was analyzed. MIN gastrocnemius mass was reduced compared to B6 mice. A 12-h fast induced MIN muscle AMPKT172 , FOXOS413 , and ULK-1S555 phosphorylation compared to B6. Wheel running attenuated these inductions. A 12-h fast induced MIN muscle MuRF-1 protein expression compared to B6 and was suppressed by wheel running. Additionally, fasting induced muscle autophagy signaling and disrupted mitochondrial quality protein expression in the MIN, which was prevented in the MIN KO. We provide evidence that increased skeletal muscle AMPK sensitivity to a 12-h fast is an adverse event in pre-cachectic MIN mice, and exercise can improve this regulation.

Exercise as a therapy for cancer-induced muscle wasting.

Cancer cachexia is a progressive disorder characterized by body weight, fat, and muscle loss. Cachexia induces metabolic disruptions that can be analogous and distinct from those observed in cancer, obscuring both diagnosis and treatment options. Inflammation, hypogonadism, and physical inactivity are widely investigated as systemic mediators of cancer-induced muscle wasting. At the cellular level, dysregulation of protein turnover and energy metabolism can negatively impact muscle mass and function. Exercise is well known for its anti-inflammatory effects and potent stimulation of anabolic signaling. Emerging evidence suggests the potential for exercise to rescue muscle's sensitivity to anabolic stimuli, reduce wasting through protein synthesis modulation, myokine release, and subsequent downregulation of proteolytic factors. To date, there is no recommendation for exercise in the management of cachexia. Given its complex nature, a multimodal approach incorporating exercise offers promising potential for cancer cachexia treatment. This review's primary objective is to summarize the growing body of research examining exercise regulation of cancer cachexia. Furthermore, we will provide evidence for exercise interactions with established systemic and cellular regulators of cancer-induced muscle wasting.

The Effect of Wheel Exercise on Functional Indices of Cachexia in Tumor-bearing Mice.

INTRODUCTION: Cancer-related fatigue and muscle wasting have received significant attention over the last few decades with the goal of establishing interventions that can improve cancer patient life quality and survival. Increased physical activity has shown to reduce cancer-associated fatigue and has been proposed as a promising therapeutic to attenuate cancer-induced wasting. However, significant gaps remain in our understanding of how physical activity affects the compositional and functional changes that initiate muscle wasting. The purpose of the current study was to determine the effect of wheel exercise on body composition and functional indices of cancer cachexia before the development of significant wasting. METHODS: Thirteen-week-old male Apc (MIN) and C57BL/6 (B6) mice were given free wheel access (W) or a locked wheel (Sed) for 5 wk. RESULTS: Wheel activity was reduced in the MIN compared with B6; however, wheel access increased complex II expression in isolated skeletal muscle mitochondria regardless of genotype. Wheel access had no effect on tumor burden or plasma interleukin-6 in the MIN. MIN-W increased body weight and lean mass compared with MIN-Sed, and there was a direct correlation between wheel distance and lean mass change. MIN-W increased grip strength and treadmill time to fatigue compared with MIN-Sed. Within MIN-W mice, skeletal muscle fatigability was only improved in high runners (>60 min·d). CONCLUSIONS: Our results suggest that there were therapeutic benefits of increased activity related to body composition, behavior, and whole-body function that were not dependent on exercise duration; however, there was an exercise threshold needed to improve skeletal muscle fatigability in tumor-bearing mice. Interestingly, wheel access was able to improve compositional and functional outcomes without mitigating tumor number or size.