Mcam stabilizes luminal progenitor breast cancer phenotypes via Ck2 control and Src/Akt/Stat3 attenuation.

Breast cancers are categorized into subtypes with distinctive therapeutic vulnerabilities and prognoses based on their expression of clinically targetable receptors and gene expression patterns mimicking different cell types of the normal gland. Here, we tested the role of Mcam in breast cancer cell state control and tumorigenicity in a luminal progenitor-like murine tumor cell line (Py230) that exhibits lineage and tumor subtype plasticity. Mcam knockdown Py230 cells show augmented Stat3 and Pi3K/Akt activation associated with a lineage state switch away from a hormone-sensing/luminal progenitor state toward alveolar and basal cell related phenotypes that were refractory to growth inhibition by the anti-estrogen therapeutic, tamoxifen. Inhibition of Stat3, or the upstream activator Ck2, reversed these cell state changes. Mcam binds Ck2 and acts as a regulator of Ck2 substrate utilization across multiple mammary tumor cell lines. In Py230 cells this activity manifests as increased mesenchymal morphology, migration, and Src/Fak/Mapk/Paxillin adhesion complex signaling in vitro, in contrast to Mcam's reported roles in promoting mesenchymal phenotypes. In vivo, Mcam knockdown reduced tumor growth and take rate and inhibited cell state transition to Sox10+/neural crest like cells previously been associated with tumor aggressiveness. This contrasts with human luminal breast cancers where MCAM copy number loss is highly coupled to Cyclin D amplification, increased proliferation, and the more aggressive Luminal B subtype. Together these data indicate a critical role for Mcam and its regulation of Ck2 in control of breast cancer cell state plasticity with implications for progression, evasion of targeted therapies and combination therapy design.

Comparison of Pain Scores and Functional Outcomes of Patients Undergoing Arthroscopic Hip Labral Repair and Concomitant Capsular Repair or Plication Versus No Closure.

Background: The need for capsular closure during arthroscopic hip labral repair is debated. Purpose: To compare pain and functional outcomes in patients undergoing arthroscopic hip labral repair with concomitant repair or plication of the capsule versus no closure. Study Design: Cohort study. Methods: Outcomes were compared between patients undergoing arthroscopic hip labral repair with concomitant repair or plication of the capsule versus no closure at up to 2 years postoperatively and with stratification by age and sex. Patients with lateral center-edge angle <20°, a history of instability, a history of prior arthroscopic surgery in the ipsilateral hip, or a history of labral debridement only were excluded. Subanalysis was performed between patients undergoing no capsular closure who were propensity score matched 1:1 with patients undergoing repair or plication based on age, sex, and preoperative Modified Harris Hip Score (MHHS). We compared patients who underwent T-capsulotomy with concomitant capsular closure matched 1:5 with patients who underwent an interportal capsulotomy with concomitant capsular repair based on age, sex, and preoperative MHHS. Results: Patients undergoing capsular closure (n = 1069), compared with the no-closure group (n = 230), were more often female (68.6% vs 53.0%, respectively; P < .001), were younger (36.4 ± 13.3 vs 47.9 ± 14.7 years; P < .001), and had superior MHHS scores at 2 years postoperatively (85.8 ± 14.5 vs 81.8 ± 18.4, respectively; P = .020). In the matched analysis, no difference was found in outcome measures between patients in the capsular closure group (n = 215) and the no-closure group (n = 215) at any follow-up timepoint. No significant difference was seen between the 2 closure techniques at any follow-up timepoint. Patients with closure of the capsule achieved the minimal clinically important difference (MCID) and the patient acceptable symptom state (PASS) for the 1-year MHHS at a similar rate as those without closure (MCID, 50.3% vs 44.9%, P = .288; PASS, 56.8% vs 51.1%, P = .287, respectively). Patients with T-capsulotomy achieved the MCID and the PASS for the 1-year MHHS at a similar rate compared with those with interportal capsulotomy (MCID, 50.1% vs 44.9%, P = .531; PASS, 65.7% vs 61.2%, P = .518, respectively). Conclusion: When sex, age, and preoperative MHHS were controlled, capsular closure and no capsular closure after arthroscopic hip labral repair were associated with similar pain and functional outcomes for patients up to 2 years postoperatively.

CRIPTO promotes extracellular vesicle uptake and activation of cancer associated fibroblasts.

Expression of CRIPTO, a factor involved in embryonic stem cells, fetal development, and wound healing, is tied to poor prognosis in multiple cancers. Prior studies in triple negative breast cancer (TNBC) models showed CRIPTO blockade inhibits tumor growth and dissemination. Here, we uncover a previously unidentified role for CRIPTO in orchestrating tumor-derived extracellular vesicle (TEV) uptake and fibroblast activation through discrete mechanisms. We found a novel mechanism by which CRIPTO drives aggressive TNBC phenotypes, involving CRIPTO-laden TEVs that program stromal fibroblasts, toward cancer associated fibroblast cell states, which in turn prompt tumor cell invasion. CRIPTO-bearing TEVs exhibited markedly elevated uptake in target fibroblasts and activated SMAD2/3 through NODAL-independent and - dependent mechanisms, respectively. Engineered expression of CRIPTO on EVs enhanced the delivery of bioactive molecules. In vivo , CRIPTO levels dictated TEV uptake in mouse lungs, a site of EV-regulated premetastatic niches important for breast cancer dissemination. These discoveries reveal a novel role for CRIPTO in coordinating heterotypic cellular crosstalk which offers novel insights into breast cancer progression, delivery of therapeutic molecules, and new, potentially targetable mechanisms of heterotypic cellular communication between tumor cells and the TME.

Safety, feasibility, and impact on the gut microbiome of kefir administration in critically ill adults.

BACKGROUND: Dysbiosis of the gut microbiome is frequent in the intensive care unit (ICU), potentially leading to a heightened risk of nosocomial infections. Enhancing the gut microbiome has been proposed as a strategic approach to mitigate potential adverse outcomes. While prior research on select probiotic supplements has not successfully shown to improve gut microbial diversity, fermented foods offer a promising alternative. In this open-label phase I safety and feasibility study, we examined the safety and feasibility of kefir as an initial step towards utilizing fermented foods to mitigate gut dysbiosis in critically ill patients. METHODS: We administered kefir in escalating doses (60 mL, followed by 120 mL after 12 h, then 240 mL daily) to 54 critically ill patients with an intact gastrointestinal tract. To evaluate kefir's safety, we monitored for gastrointestinal symptoms. Feasibility was determined by whether patients received a minimum of 75% of their assigned kefir doses. To assess changes in the gut microbiome composition following kefir administration, we collected two stool samples from 13 patients: one within 72 h of admission to the ICU and another at least 72 h after the first stool sample. RESULTS: After administering kefir, none of the 54 critically ill patients exhibited signs of kefir-related bacteremia. No side effects like bloating, vomiting, or aspiration were noted, except for diarrhea in two patients concurrently on laxatives. Out of the 393 kefir doses prescribed for all participants, 359 (91%) were successfully administered. We were able to collect an initial stool sample from 29 (54%) patients and a follow-up sample from 13 (24%) patients. Analysis of the 26 paired samples revealed no increase in gut microbial α-diversity between the two timepoints. However, there was a significant improvement in the Gut Microbiome Wellness Index (GMWI) by the second timepoint (P = 0.034, one-sided Wilcoxon signed-rank test); this finding supports our hypothesis that kefir administration can improve gut health in critically ill patients. Additionally, the known microbial species in kefir were found to exhibit varying levels of engraftment in patients' guts. CONCLUSIONS: Providing kefir to critically ill individuals is safe and feasible. Our findings warrant a larger evaluation of kefir's safety, tolerability, and impact on gut microbiome dysbiosis in patients admitted to the ICU. TRIAL REGISTRATION: NCT05416814; trial registered on June 13, 2022.

Pain scores and functional outcomes of patients with shoulder labral repair using all-suture anchors versus conventional anchors.

HYPOTHESIS/PURPOSE: The purpose of this study was to compare PROMs in patients undergoing anterior glenoid labral repair using all-suture versus conventional anchors. We hypothesized PROMs would be similar between groups. METHODS: We performed a retrospective review of the Arthrex Global Surgical Outcomes System (SOS) database, querying patients who underwent arthroscopic glenoid labral repair between 01/01/2015 and 12/31/2020. Patients aged 18-100, who had isolated glenoid labrum repair with at least 12-month follow-up were included. The visual analog pain scale (VAS), Western Ontario Shoulder Instability Index, Veteran's RAND 12-items health survey, single assessment numeric evaluation and the American Shoulder and Elbow Surgeons score (ASES) were compared preoperatively, 3 months, 6 months, 1 year and 2 years postoperatively in patients who received all-suture anchors versus conventional anchors in the setting of anterior glenoid labrum repair. Our primary aim was comparison of PROMs between patients receiving all-suture versus conventional suture anchors. Secondarily, a sub-analysis was performed comparing outcomes based on anchor utilization for patients with noted anterior instability. RESULTS: We evaluated 566 patients, 54 patients receiving all-suture anchors and 512 patients receiving conventional anchors. At two-year follow-up there was no significant difference between the two groups in PROMs. In a sub-analysis of isolated anterior labrum repair, there was an improvement in ASES (P = 0.034) and VAS (P = 0.039) with the all-suture anchor at two-year follow-up. CONCLUSIONS: All-suture anchors provide similar or superior pain and functional outcome scores up to 2 years postoperatively compared to conventional anchors. CLINICAL RELEVANCE: As all-suture anchors gain popularity among surgeons, this is the largest scale study to date validating their use in the setting of glenoid labrum repair. Institutional Review Board (IRB): IRB202102550.

Endoscope-assisted three-wire technique for extensive nasal septum resection in horses.

OBJECTIVE: To describe a 3-wire method with endoscopic guidance for extensive nasal septum resection. STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Thirteen horses with nasal septum diseases. METHODS: In anesthetized horses in left lateral recumbency, endoscopic guidance was used to place obstetrical wires for the ventral and caudal incisions in the nasal septum and a trephine opening was used to place the dorsal wire. The rostral aspect of the septum was incised with a scalpel, followed by incisions with the preplaced wires, and the nasal passages were packed with gauze. Horses were recovered with a temporary tracheotomy. RESULTS: Conversion to intraoral placement of wires was required in two horses, one to correct entangled wires and the other because hemorrhage obscured the endoscopic view. Exercise tolerance was improved postoperatively, abnormal respiratory noise was decreased or eliminated by surgery in all horses, and all owners were satisfied. One Thoroughbred racehorse performed with modest success. CONCLUSIONS: Modification of the 3-wire method was effective and safe for extensive nasal septum removal. Technical complications of the procedure include entangling of wires and intraoperative hemorrhage. CLINICAL SIGNIFICANCE: Endoscopic guidance can be used to place obstetrical wires for nasal septum resection in small horses and precludes use of a large tracheotomy for anesthetic delivery. Reasons for athletic failures were difficult to establish retrospectively, although assessment of postoperative noise at speed might be more relevant to recovery of athletic potential than assessment at slower gaits.

Nonsteroidal antiinflammatory drugs reduce anion secretion and increase apoptosis in equine colonic mucosa ex vivo.

OBJECTIVE: Right dorsal colitis causes chronic colic associated with long-term treatment with nonsteroidal antiinflammatory drugs (NSAIDs). This study was designed to determine if NSAIDs could inhibit anion transporters that protect against intestinal mucosal injury in other species. ANIMALS: 20 healthy horses. METHODS: The effects of indomethacin (INDO) and firocoxib (FIR), on short-circuit current (Isc) in mucosa from the right dorsal colon (RDC) and right ventral colon (RVC) were measured in Ussing chambers by standard electrophysiological techniques. Immunohistochemical methods were used to detect apoptosis (caspase-3) with these NSAIDs and phenylbutazone (PBZ) and to locate the NKCC1 transporter. RESULTS: The Isc in RDC and RVC incubated with INDO or FIR was increased almost 3-fold (P < .0001) by prostaglandin E2 (PGE2) through a system inhibited by loop diuretics (P < .0001). Although these findings and anion replacement studies were consistent with anion secretion, the RDC also displayed an Isc response suggestive of a unique transporter apparently absent in RVC or NSAID-free solutions. In RDC, FIR, INDO, and PBZ induced apoptosis in the lower half of crypts. However, significant differences in apoptotic index were recorded in the RDC between NSAID-treated and control tissues (no NSAID). CLINICAL RELEVANCE: The effects of NSAIDs on Isc were consistent with reduced anion secretion, which could represent the pharmacological equivalent of the transport failure responsible for Cystic Fibrosis (CF) in other species. Failure of anion secretion could interfere with buffering acid from intraluminal fermentation, which could suggest a treatment target for right dorsal colitis.

Discovery of KB-0742, a Potent, Selective, Orally Bioavailable Small Molecule Inhibitor of CDK9 for MYC-Dependent Cancers.

Transcriptional deregulation is a hallmark of many cancers and is exemplified by genomic amplifications of the MYC family of oncogenes, which occur in at least 20% of all solid tumors in adults. Targeting of transcriptional cofactors and the transcriptional cyclin-dependent kinase (CDK9) has emerged as a therapeutic strategy to interdict deregulated transcriptional activity including oncogenic MYC. Here, we report the structural optimization of a small molecule microarray hit, prioritizing maintenance of CDK9 selectivity while improving on-target potency and overall physicochemical and pharmacokinetic (PK) properties. This led to the discovery of the potent, selective, orally bioavailable CDK9 inhibitor 28 (KB-0742). Compound 28 exhibits in vivo antitumor activity in mouse xenograft models and a projected human PK profile anticipated to enable efficacious oral dosing. Notably, 28 is currently being investigated in a phase 1/2 dose escalation and expansion clinical trial in patients with relapsed or refractory solid tumors.

Repeat Celiotomy-Current Status.

Repeat celiotomy can be lifesaving in horses with a surgically treatable postoperative obstruction, although guidelines for its use are lacking, except for uncontrollable postoperative pain. Overdiagnosis of ileus as the cause of postoperative obstruction could delay a second surgery so the disease progresses beyond a manageable level of severity. Although many horses respond favorably to repeat celiotomy, complications can be severe and life threatening, such as incisional infection and adhesions. Repeat celiotomy does not seem to exacerbate postoperative ileus, despite additional surgical manipulation. An important benefit of repeat celiotomy is termination of hopeless cases, thereby reducing cost and suffering.

Surgical enlargement of the epiploic foramen in horses.

OBJECTIVE: To assess the safety and efficacy of a method for digitally enlarging the caudal aspect of the epiploic foramen (EF). STUDY DESIGN: Healthy horses and clinical cases of EF entrapment (EFE). ANIMALS: Fourteen healthy horses and three clinical cases. METHODS: Through a ventral midline celiotomy under general anesthesia, the EF was enlarged by digital separation of the caudal attachments of the caudate lobe of the liver from right dorsal colon, right kidney, gastropancreatic fold, and pancreas. Healthy horses were euthanized under anesthesia, and the enlarged EF was measured at necropsy. RESULTS: The method used for enlarging the EF did not cause clinically relevant hemorrhage, as determined by visual inspection of the EF in 14 horses at necropsy and by vital parameters under anesthesia in all horses. In clinical cases, EFE was reduced following enlargement of the EF, and no intraoperative complications were encountered. In one clinical case, necropsy at 30 days confirmed partial closure of the enlarged EF. CONCLUSION: The method proposed enlarged the EF safely and effectively. Limitations of the study include the small number of clinical cases and the lack of postoperative follow-up on the healthy horses. CLINICAL SIGNIFICANCE: Enlargement of the EF at its caudal extent should be considered in selected cases of EFE in which manual reduction is difficult or protracted. Although the procedure was safe in this study, knowledge of the anatomy, practice on cadavers, and careful selection of cases with greatest need are recommended before clinical use.

Ex vivo and in vivo evaluation of a modified interrupted Lembert pattern for small intestinal anastomoses in horses.

OBJECTIVE: To compare single-layer anastomoses (modified continuous Lembert [mod-CL] and modified-interrupted Lembert [mod-IL]) and a 2-layer simple continuous anastomosis (2 L; seromuscular and mucosal) for jejunojejunal anastomoses in equine cadavers and to compare ex vivo to in vivo time to complete the anastomosis and stoma size with a mod-IL pattern. STUDY DESIGN: Measurements in jejunum from cadaver and anesthetized horses. ANIMALS: Ten live horses and 18 equine cadavers. METHODS: Time to complete anastomosis, bursting pressures (BP), leakage sites, and anastomotic index (size ratio of anastomotic lumen to control lumen) were recorded. Time to completion and lumen size were compared between in vivo and ex vivo mod-IL patterns. RESULTS: The mod-CL pattern was fastest (8.44 ± 1.30 min, p < .05), and the 2 L pattern was slower (17.07 ± 2.0 min) than the mod-CL and mod-IL (p < .05). The anastomotic index exceeded 100 and did not differ between patterns. Segments reached higher bursting pressures when anastomosed with mod-IL (145.94 ± 24.18 mm Hg) than mod-CL (p < .05). In vivo closure was approximately 8 minutes slower than ex vivo, and with a smaller anastomotic index. CONCLUSIONS: All anastomoses increased lumen size over control segments ex vivo. Lumen size after placement of a mod-IL was greater ex vivo than in vivo, and completion was slower in vivo than ex vivo. [Corrections added on 26 Dec 2022, after online publication: added "ex vivo" to the first line of Conclusions in the Abstract.] CLINICAL SIGNIFICANCE: Slower and smaller anastomoses should be anticipated in vivo compared to ex vivo results. Anastomoses with a mod-IL pattern appear clinically advantageous, producing a comparable lumen size in less time than 2 L.

Not just a cousin of the naked mole-rat: Damaraland mole-rats offer unique insights into biomedicine.

Evolutionary medicine has been a fast-growing field of biological research in the past decade. One of the strengths of evolutionary medicine is to use non-traditional model organisms which often exhibit unusual characteristics shaped by natural selection. Studying these unusual traits could provide valuable insight to understand biomedical questions, since natural selection likely discovers solutions to those complex biological problems. Because of many unusual traits, the naked mole-rat (NMR) has attracted attention from different research areas such as aging, cancer, and hypoxia- and hypercapnia-related disorders. However, such uniqueness of NMR physiology may sometimes make the translational study to human research difficult. Damaraland mole-rat (DMR) shares multiple characteristics in common with NMR, but shows higher degree of similarity with human in some aspects of their physiology. Research on DMR could therefore offer alternative insights and might bridge the gap between experimental findings from NMR to human biomedical research. In this review, we discuss studies of DMR as an extension of the current set of model organisms to help better understand different aspects of human biology and disease. We hope to encourage researchers to consider studying DMR together with NMR. By studying these two similar but evolutionarily distinct species, we can harvest the power of convergent evolution and avoid the potential biased conclusions based on life-history of a single species.

Effects of age, disease and anastomosis on short- and long-term survival after surgical correction of small intestinal strangulating diseases in 89 horses.

BACKGROUND: Although survival rates have been reported after small intestinal surgery for strangulating diseases in horses, none have followed survival for periods relevant to the long lifespan of horses and none have described effect of age, disease and surgical treatments over such long survival periods. OBJECTIVES: To examine effects of age, disease and type of surgery on long-term survival in horses after surgical treatment of small intestinal strangulating diseases over periods relevant to the expected lifespan of a horse. STUDY DESIGN: Retrospective clinical study. METHODS: Post-operative data were gathered from medical records and owner contact for 89 horses with small intestinal strangulation. Survival times from surgery to the date of death or the date of last follow-up were analysed by Kaplan-Meier statistics. Variables of interest were age, type of strangulating disease and surgical correction. Cox proportional hazards regression was used to evaluate these variables. RESULTS: Short-term survival was not affected by any of the variables measured. For long-term survival with Kaplan-Meier statistics, horses ≥16 years old had significantly shorter (P = .002) median survival times (72 months; 95% CI 32.0-96.0) than younger horses (121.7 months; 95% CI 90.0-162), horses without resection had significantly longer (P = .02) survival times (120 months; 95% CI 86-212) than horses that had jejunocecostomy (76.8 months; 95% CI 24-125), and horses with miscellaneous diseases had significantly longer (P = .02) median survival times (161.9 months; 95% CI 72.0-M) than horses with strangulating lipoma (79.8 months; 95% CI 32.0-120.0). In the multivariable Cox Proportional Hazard model, age (HR = 2.67; 1.49-4.75, P < .001) and anastomosis (HR = 0.65; 0.46-0.92, P = .02) had the most significant effect on median survival time. MAIN LIMITATIONS: Limitations were small numbers in some categories, loss of cases to follow-up, owner recall failures and lack of a control group. CONCLUSIONS: The remaining lifespan of older horses at the time of surgery had the greatest effect on survival. Age could influence long-term survival studies after colic surgery, and therefore needs to be considered for survival analyses. Horses that did not require resection and anastomosis had favourable outcomes, underscoring the potential importance of early intervention to reduce the need for resection.

Ventral cervico-thoracic meningeal cyst resulting in myelopathy: Case report and literature review.

BACKGROUND: Spinal meningeal (dural) cysts rarely cause spinal cord compression and/or myelopathy. CASE DESCRIPTION: A 38-year-old male presented with 6 weeks of worsening bilateral lower extremity paresthesias and an unsteady gait. Notably, the patient was involved in a snowmobile accident 7 years ago that resulted in trauma to his thoracic spine for which he had undergone a corpectomy and posterior fusion. A full spine MRI was obtained to evaluate his new paresthesias and myelopathy, which revealed a large extra-axial fluid collection consistent with a meningeal cyst extending from C2 to T4. This caused severe spinal cord compression, maximal at the T1-3 level. The patient underwent a T1-3 laminectomy initially accompanied by partial cyst resection/ drainage, but ultimately he returned and required a subsequent cystoperitoneal shunt. Following the final surgery, the patient's symptoms gradually resolved over 6 months postoperatively. CONCLUSION: Spinal meningeal cysts rarely cause back pain and/or neurological symptoms. MRI is the diagnostic study of choice for defining this entity. Operative intervention must be tailored to the symptoms, location, extent, and type of the cyst. If cysts recur after partial resection and drainage, cystoperitoneal shunt placement is warranted.

Whence CRIPTO: The Reemergence of an Oncofetal Factor in 'Wounds' That Fail to Heal.

There exists a set of factors termed oncofetal proteins that play key roles in ontogeny before they decline or disappear as the organism's tissues achieve homeostasis, only to then re-emerge in cancer. Although the unique therapeutic potential presented by such factors has been recognized for more than a century, their clinical utility has yet to be fully realized1. This review highlights the small signaling protein CRIPTO encoded by the tumor derived growth factor 1 (TDGF1/Tdgf1) gene, an oft cited oncofetal protein whose presence in the cancer literature as a tumor promoter, diagnostic marker and viable therapeutic target continues to grow. We touch lightly on features well established and well-reviewed since its discovery more than 30 years ago, including CRIPTO's early developmental roles and modulation of SMAD2/3 activation by a selected set of transforming growth factor ß (TGF-ß) family ligands. We predominantly focus instead on more recent and less well understood additions to the CRIPTO signaling repertoire, on its potential upstream regulators and on new conceptual ground for understanding its mode of action in the multicellular and often stressful contexts of neoplastic transformation and progression. We ask whence it re-emerges in cancer and where it 'hides' between the time of its fetal activity and its oncogenic reemergence. In this regard, we examine CRIPTO's restriction to rare cells in the adult, its potential for paracrine crosstalk, and its emerging role in inflammation and tissue regeneration-roles it may reprise in tumorigenesis, acting on subsets of tumor cells to foster cancer initiation and progression. We also consider critical gaps in knowledge and resources that stand between the recent, exciting momentum in the CRIPTO field and highly actionable CRIPTO manipulation for cancer therapy and beyond.

J-incision to approach the cranial abdomen in the adult horse.

OBJECTIVE: To describe a modified celiotomy to improve access to cranial abdominal structures in horses. ANIMALS: Four horses. STUDY DESIGN: Short case series. METHODS: Three horses with gastric impactions were treated with gastrotomies. One horse was treated for a diaphragmatic hernia with herniorrhaphy and mesh augmentation. In all horses, the ventral midline celiotomy was modified cranially with a J-incision through the body wall, along the paracostal arch. RESULTS: The only surgical complications were midline incisional infections in all horses. Three of the four horses had good long-term outcomes; the remaining horse underwent euthanasia for reasons likely unrelated to incisional complications. CONCLUSION: The J-incision improved access to the stomach and diaphragm in these horses. The paracostal component healed in all cases without evidence of infection or dehiscence. CLINICAL SIGNIFICANCE: This modified celiotomy may be considered to improve access during gastrotomy and repair of dorsally located diaphragmatic hernias.

Modulating Androgen Receptor-Driven Transcription in Prostate Cancer with Selective CDK9 Inhibitors.

Castration-resistant prostate cancers (CRPCs) lose sensitivity to androgen-deprivation therapies but frequently remain dependent on oncogenic transcription driven by the androgen receptor (AR) and its splice variants. To discover modulators of AR-variant activity, we used a lysate-based small-molecule microarray assay and identified KI-ARv-03 as an AR-variant complex binder that reduces AR-driven transcription and proliferation in prostate cancer cells. We deduced KI-ARv-03 to be a potent, selective inhibitor of CDK9, an important cofactor for AR, MYC, and other oncogenic transcription factors. Further optimization resulted in KB-0742, an orally bioavailable, selective CDK9 inhibitor with potent anti-tumor activity in CRPC models. In 22Rv1 cells, KB-0742 rapidly downregulates nascent transcription, preferentially depleting short half-life transcripts and AR-driven oncogenic programs. In vivo, oral administration of KB-0742 significantly reduced tumor growth in CRPC, supporting CDK9 inhibition as a promising therapeutic strategy to target AR dependence in CRPC.

CRIPTO antagonist ALK4L75A-Fc inhibits breast cancer cell plasticity and adaptation to stress.

BACKGROUND: CRIPTO is a multi-functional signaling protein that promotes stemness and oncogenesis. We previously developed a CRIPTO antagonist, ALK4L75A-Fc, and showed that it causes loss of the stem cell phenotype in normal mammary epithelia suggesting it may similarly inhibit CRIPTO-dependent plasticity in breast cancer cells. METHODS: We focused on two triple negative breast cancer cell lines (MDA-MB-231 and MDA-MB-468) to measure the effects of ALK4L75A-Fc on cancer cell behavior under nutrient deprivation and endoplasmic reticulum stress. We characterized the proliferation and migration of these cells in vitro using time-lapse microscopy and characterized stress-dependent changes in the levels and distribution of CRIPTO signaling mediators and cancer stem cell markers. We also assessed the effects of ALK4L75A-Fc on proliferation, EMT, and stem cell markers in vivo as well as on tumor growth and metastasis using inducible lentiviral delivery or systemic administration of purified ALK4L75A-Fc, which represents a candidate therapeutic approach. RESULTS: ALK4L75A-Fc inhibited adaptive responses of breast cancer cells under conditions of nutrient and ER stress and reduced their proliferation, migration, clonogenicity, and expression of EMT and cancer stem cell markers. ALK4L75A-Fc also inhibited proliferation of human breast cancer cells in stressed tumor microenvironments in xenografts and reduced both primary tumor size and metastatic burden. CONCLUSIONS: Cancer cell adaptation to stresses such as nutrient deprivation, hypoxia, and chemotherapy can critically contribute to dormancy, metastasis, therapy resistance, and recurrence. Identifying mechanisms that govern cellular adaptation, plasticity, and the emergence of stem-like cancer cells may be key to effective anticancer therapies. Results presented here indicate that targeting CRIPTO with ALK4L75A-Fc may have potential as such a therapy since it inhibits breast cancer cell adaptation to microenvironmental challenges and associated stem-like and EMT phenotypes.

Association between GDF15, poverty and mortality in urban middle-aged African American and white adults.

Mortality disparities are influenced by race and poverty. There is limited information about whether poverty influences biologic markers of mortality risk. Emerging data suggests that growth differentiation factor 15 (GDF15) is associated with mortality; however, the interplay between GDF15, sociodemographic factors and mortality is not known. We sought to evaluate the interactions between GDF15 and sex, race and poverty status on mortality. Serum GDF15 was measured in 1036 African American and white middle-aged men and women above and below 125% of the Federal poverty status from the Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) study. Multivariable adjusted Cox regression models were used to assess the association between log-transformed GDF15 (logGDF15) and 12-year mortality outcomes (all-cause, cardiovascular- and cancer-specific outcomes) and interactions with sex, race and poverty status. Likelihood ratio tests were used to assess significance of the interaction terms. Median GDF15 was 655.2 pg/mL (IQR = 575.1). During 12.2 years of follow-up, 331 died of which 94 cardiovascular- and 87 were cancer-specific deaths. One unit of increase in logGDF15 was associated with a hazard ratio for all-cause mortality, cardiovascular- and cancer-specific mortality of 2.26 (95% confidence interval [CI], 1.94-2.64), 2.74 (95%CI, 2.06-3.63) and 1.41 (95%CI, 1.00-2.00), respectively. There was an interaction between logGDF15 and poverty status on all-cause mortality (p<0.05). The GDF15×poverty status interaction term improved model calibration for all-cause mortality. Our study provides the first evidence that the effect of elevated GDF15 on all-cause mortality is modified by poverty status.