Search

Background: Meniscal deficiency has been reported to increase contact pressures in the affected tibiofemoral joint, possibly leading to degenerative changes. Current surgical options include meniscal allograft transplantation and insertion of segmental meniscal scaffolds. Little is known about segmental meniscal allograft transplantation. Purpose: To evaluate the effectiveness of segmental medial meniscal allograft transplantation in the setting of partial medial meniscectomy in restoring native knee loading characteristics. Study Design: Controlled laboratory study. Methods: Ten fresh-frozen human cadaveric knees underwent central midbody medial meniscectomy and subsequent segmental medial meniscal allograft transplantation. Knees were loaded in a dynamic tensile testing machine to 1000 N for 20 seconds at 0°, 30°, 60°, and 90° of flexion. Four conditions were tested: (1) intact medial meniscus, (2) deficient medial meniscus, (3) segmental medial meniscal transplant fixed with 7 meniscocapsular sutures, and (4) segmental medial meniscal transplant fixed with 7 meniscocapsular sutures and 1 suture fixed through 2 bone tunnels. Submeniscal medial and lateral pressure-mapping sensors assessed mean contact pressure, peak contact pressure, mean contact area, and pressure mapping. Two-factor random-intercepts linear mixed effects models compared pressure and contact area measurements among experimental conditions. Results: The meniscal-deficient state demonstrated a significantly higher mean contact pressure than all other testing conditions (mean difference, ≥0.35 MPa; P < .001 for all comparisons) and a significantly smaller total contact area as compared with all other testing conditions (mean difference, ≤140 mm2; P < .001 for all comparisons). There were no significant differences in mean contact pressure or total contact area among the intact, transplant, or transplant-with-tunnel groups or in any outcome measure across all comparisons in the lateral compartment. No significant differences existed in center of pressure and relative pressure distribution across testing conditions. Conclusion: Segmental medial meniscal allograft transplantation restored the medial compartment mean contact pressure and mean contact area to values measured in the intact medial compartment. Clinical Relevance: Segmental medial meniscal transplantation may provide an alternative to full meniscal transplantation by addressing only the deficient portion of the meniscus with transplanted tissue. Additional work is required to validate long-term fixation strength and biologic integration.

PURPOSE: Therapeutic efficacy of hormonal therapies to target estrogen receptor (ER)-positive breast cancer is limited by the acquisition of ligand-independent ESR1 mutations, which confer treatment resistance to aromatase inhibitors (AIs). Monitoring for the emergence of such mutations may enable individualized therapy. We thus assessed CTC- and ctDNA-based detection of ESR1 mutations with the aim of evaluating non-invasive approaches for the determination of endocrine resistance. PATIENTS AND METHODS: In a prospective cohort of 55 women with hormone receptor-positive metastatic breast cancer, we isolated circulating tumor cells (CTCs) and developed a high-sensitivity method for the detection of ESR1 mutations in these CTCs. In patients with sufficient plasma for the simultaneous extraction of circulating tumor DNA (ctDNA), we performed a parallel analysis of ESR1 mutations using multiplex droplet digital PCR (ddPCR) and examined the agreement between these two platforms. Finally, we isolated single CTCs from a subset of these patients and reviewed RNA expression to explore alternate methods of evaluating endocrine responsiveness. RESULTS: High-sensitivity ESR1 sequencing from CTCs revealed mono- and oligoclonal mutations in 22% of patients. These were concordant with plasma DNA sequencing in 95% of cases. Emergence of ESR1 mutations was correlated both with time to metastatic relapse and duration of AI therapy following such recurrence. The Presence of an ESR1 mutation, compared to ESR1 wild type, was associated with markedly shorter Progression-Free Survival on AI-based therapies (p = 0.0006), but unaltered to other non-AI-based therapies (p = 0.73). Compared with ESR1 mutant cases, AI-resistant CTCs with wild-type ESR1 showed an elevated ER-coactivator RNA signature, consistent with their predicted response to second-line hormonal therapies. CONCLUSION: Blood-based serial monitoring may guide the selection of precision therapeutics for women with AI-resistant ER-positive breast cancer.

Meniscal tears treated with partial meniscectomies have been shown to significantly increase contract pressures within the tibiofemoral joint, and a complete focal meniscal deficiency may render the entirety of the meniscus functionally incompetent. Although various techniques of meniscal transplantation have been described, these techniques may require the excision of a considerable amount of healthy meniscal tissue. Furthermore, failures continue to frequently occur. Therefore, attempts to restoring normal knee kinematics and biomechanical forces are essential. Segmental meniscus allograft transplantations may offer the advantage of a robust repair by both maintaining knee biomechanics and biology while maximizing preservation of native meniscal tissue. Also, most meniscal deficiency involves only a portion of the meniscus, and thus we developed this technique to segmentally transplant only the deficient portion. The purpose of this Technical Note is to describe a technique of segmental medial meniscus allograft transplantation in a patient with focal medial meniscus deficiency., Technique Video Video 1 This video outlines the step-by-step illustration of anterior cruciate ligament reconstruction and segmental medial meniscus transplant in a patient with focal medial meniscus deficiency. We perform this technique by first performing a diagnostic arthroscopy, which reveals the region of full-thickness segmental meniscus deficiency. After debridement, a spinal needle is passed from outside-in to determine the location of the defect and to pass a shuttling suture to eventually shuttle the prepared meniscus transplant into the defect. Next, the meniscus allograft is prepared by marking both the posterior and anterior aspects, and the desired length of transplant is measured. The 20-mm allograft of meniscus is excised from the donor meniscus. Preparation of the graft consists of No. 2 nonabsorbable sutures around the periphery of the graft. A total of 5 sutures are passed around the periphery in a vertical mattress configuration. A midline incision and a medial parapatellar arthrotomy are performed to accomplish both the proximal tibia osteotomy and the 2 osteochondral allograft plugs. The same arthrotomy is used to shuttle the meniscus graft. Once the allograft is positioned intraarticularly, the 5 vertical mattress sutures are passed through the capsule using a needle-shuttling device. The anterior and posterior regions of the graft are then incorporated into native tissue using No. 2 nonabsorbable sutures. The transplant is further anchored anteriorly to the tibia with a single 2.9-mm suture anchor placed at the periphery of the tibia. The final construct was intact to probing and stable.

Blood-borne metastasis to the brain is a major complication of breast cancer, but cellular pathways that enable cancer cells to selectively grow in the brain microenvironment are poorly understood. We find that cultured circulating tumor cells (CTCs), derived from blood samples of women with advanced breast cancer and directly inoculated into the mouse frontal lobe, exhibit striking differences in proliferative potential in the brain. Derivative cell lines generated by serial intracranial injections acquire selectively increased proliferative competency in the brain, with reduced orthotopic tumor growth. Increased Hypoxia Inducible Factor 1A (HIF1A)-associated signaling correlates with enhanced proliferation in the brain, and shRNA-mediated suppression of HIF1A or drug inhibition of HIF-associated glycolytic pathways selectively impairs brain tumor growth while minimally impacting mammary tumor growth. In clinical specimens, brain metastases have elevated HIF1A protein expression, compared with matched primary breast tumors, and in patients with brain metastases, hypoxic signaling within CTCs predicts decreased overall survival. The selective activation of hypoxic signaling by metastatic breast cancer in the brain may have therapeutic implications., The mechanisms underlying the growth of breast cancer metastasis in the brain are unclear. Here, the authors use an intracranial injection mouse model and single-cell analysis of patient circulating tumour cells to demonstrate that increased hypoxic and HIF1A signalling promotes tumour growth in the brain.

Purpose: Plasma genotyping may identify mutations in potentially “actionable” cancer genes, such as BRCA1/2, but their clinical significance is not well-defined. We evaluated the characteristics of somatically acquired BRCA1/2 mutations in patients with metastatic breast cancer (MBC). Experimental Design: Patients with MBC undergoing routine cell-free DNA (cfDNA) next-generation sequencing (73-gene panel) before starting a new therapy were included. Somatic BRCA1/2 mutations were classified as known germline pathogenic mutations or novel variants, and linked to clinicopathologic characteristics. The effect of the PARP inhibitor, olaparib, was assessed in vitro, using cultured circulating tumor cells (CTCs) from a patient with a somatically acquired BRCA1 mutation and a second patient with an acquired BRCA2 mutation. Results: Among 215 patients with MBC, 29 (13.5%) had somatic cfDNA BRCA1/2 mutations [nine (4%) known germline pathogenic and rest (9%) novel variants]. Known germline pathogenic BRCA1/2 mutations were common in younger patients (P = 0.008), those with triple-negative disease (P = 0.022), and they were more likely to be protein-truncating alterations and be associated with TP53 mutations. Functional analysis of a CTC culture harboring a somatic BRCA1 mutation demonstrated high sensitivity to PARP inhibition, while another CTC culture harboring a somatic BRCA2 mutation showed no differential sensitivity. Across the entire cohort, APOBEC mutational signatures (COSMIC Signatures 2 and 13) and the “BRCA” mutational signature (COSMIC Signature 3) were present in BRCA1/2-mutant and wild-type cases, demonstrating the high mutational burden associated with advanced MBC. Conclusions: Somatic BRCA1/2 mutations are readily detectable in MBC by cfDNA analysis, and may be present as both known germline pathogenic and novel variants.

To report patient characteristics, fracture types, treatment methods, early clinical outcomes and complications of children and adolescents treated for tibial tubercle fractures. Retrospective case series of patients 18 years old and younger treated for tibial tubercle fractures at a single institution from 1995 to 2015. Clinical and radiographic outcomes were reported at minimum six-month follow-up. In 228 patients, 236 tibial tubercle fractures were identified, of whom, 198 (87%) were males. Mean age and BMI was 14.3 years and 25.0, respectively. Pre-existing Osgood-Schlatter disease was identified in 31% cases and was most commonly associated with type I fractures (P < 0.001). Most fractures occurred while participating in athletics (86%). Initial treatment was surgical for 67% fractures. Type III fractures were most common (41%), followed by type I (29%). Type I fractures were most commonly treated nonoperatively (91%) and types II-V fractures were most commonly treated surgically (89%, P < 0.001). Compartment syndrome was identified in 4 (2%) patients, 3 of which had type IV fractures. Most patients returned to sports (88%). Tibial tubercle fractures are sports-related injuries and occur most commonly in adolescent males. They can be associated with pre-existing Osgood-Schlatter disease, a higher than average BMI, and a small but relevant risk of compartment syndrome. Following treatment, most patients return to sports participation.

Long head of the biceps (LHB) pathology is a prevalent cause of shoulder pain. Arthroscopic tenotomy and tenodesis are performed for treatment at increasing frequency. When LHB pathology is the only glenohumeral intra-articular pathology that needs to be addressed, and an LHB tenotomy or subpectoral LHB tenodesis is planned, it is unnecessary and potentially harmful to establish an anterior rotator interval portal. The objective of this Technical Note is to describe a minimally invasive technique for LHB tenotomy at the supraglenoid tubercle without the need for establishing an accessory portal.

Radiologic screening of high-risk adults reduces lung-cancer-related mortality1,2; however, a small minority of eligible individuals undergo such screening in the United States3,4. The availability of blood-based tests could increase screening uptake. Here we introduce improvements to cancer personalized profiling by deep sequencing (CAPP-Seq)5, a method for the analysis of circulating tumour DNA (ctDNA), to better facilitate screening applications. We show that, although levels are very low in early-stage lung cancers, ctDNA is present prior to treatment in most patients and its presence is strongly prognostic. We also find that the majority of somatic mutations in the cell-free DNA (cfDNA) of patients with lung cancer and of risk-matched controls reflect clonal haematopoiesis and are non-recurrent. Compared with tumour-derived mutations, clonal haematopoiesis mutations occur on longer cfDNA fragments and lack mutational signatures that are associated with tobacco smoking. Integrating these findings with other molecular features, we develop and prospectively validate a machine-learning method termed 'lung cancer likelihood in plasma' (Lung-CLiP), which can robustly discriminate early-stage lung cancer patients from risk-matched controls. This approach achieves performance similar to that of tumour-informed ctDNA detection and enables tuning of assay specificity in order to facilitate distinct clinical applications. Our findings establish the potential of cfDNA for lung cancer screening and highlight the importance of risk-matching cases and controls in cfDNA-based screening studies.

Small-cell lung cancer (SCLC) is an aggressive malignancy in which inhibitors of PARP have modest single-agent activity. We performed a phase I/II trial of combination olaparib tablets and temozolomide (OT) in patients with previously treated SCLC. We established a recommended phase II dose of olaparib 200 mg orally twice daily with temozolomide 75 mg/m2 daily, both on days 1 to 7 of a 21-day cycle, and expanded to a total of 50 patients. The confirmed overall response rate was 41.7% (20/48 evaluable); median progression-free survival was 4.2 months [95% confidence interval (CI), 2.8–5.7]; and median overall survival was 8.5 months (95% CI, 5.1–11.3). Patient-derived xenografts (PDX) from trial patients recapitulated clinical OT responses, enabling a 32-PDX coclinical trial. This revealed a correlation between low basal expression of inflammatory-response genes and cross-resistance to both OT and standard first-line chemotherapy (etoposide/platinum). These results demonstrate a promising new therapeutic strategy in SCLC and uncover a molecular signature of those tumors most likely to respond. Significance: We demonstrate substantial clinical activity of combination olaparib/temozolomide in relapsed SCLC, revealing a promising new therapeutic strategy for this highly recalcitrant malignancy. Through an integrated coclinical trial in PDXs, we then identify a molecular signature predictive of response to OT, and describe the common molecular features of cross-resistant SCLC. See related commentary by Pacheco and Byers, p. 1340. This article is highlighted in the In This Issue feature, p. 1325

Plasma genotyping identifies potentially actionable mutations at variable mutant allele frequencies, often admixed with multiple subclonal variants, highlighting the need for their clinical and functional validation. We prospectively monitored plasma genotypes in 143 women with endocrine-resistant metastatic breast cancer (MBC), identifying multiple novel mutations including HER2 mutations (8.4%), albeit at different frequencies highlighting clinical heterogeneity. To evaluate functional significance, we established ex vivo culture from circulating tumor cells (CTCs) from a patient with HER2-mutant MBC, which revealed resistance to multiple targeted therapies including endocrine and CDK 4/6 inhibitors, but high sensitivity to neratinib (IC50: 0.018 μM). Immunoblotting analysis of the HER2-mutant CTC culture line revealed high levels of HER2 expression at baseline were suppressed by neratinib, which also abrogated downstream signaling, highlighting oncogenic dependency with HER2 mutation. Furthermore, treatment of an index patient with HER2-mutant MBC with the irreversible HER2 inhibitor neratinib resulted in significant clinical response, with complete molecular resolution of two distinct clonal HER2 mutations, with persistence of other passenger subclones, confirming HER2 alteration as a driver mutation. Thus, driver HER2 mutant alleles that emerge during blood-based monitoring of endocrine-resistant MBC confer novel therapeutic vulnerability, and ex vivo expansion of viable CTCs from the blood circulation may broadly complement plasma-based mutational analysis in MBC.

Background and purpose Distal biceps rupture is less common than injury to the proximal biceps; however, injury distally has profound functional implications on activities which rely on power during elbow flexion and forearm supination. The majority of distal biceps ruptures can be treated with surgical repair of the distal biceps utilizing either a single or two-incision technique; both of which achieve comparable improved outcomes and reported minimal pain and disability at two years. Safe and effective rehabilitation following distal biceps repair is accomplished through a phased progression, with avoidance of premature stress to the healing soft tissue repair.The purpose of this clinical commentary is to provide a concise review of distal biceps tendon injury, including relevant anatomy, etiology, diagnosis, and operative intervention as well as post-operative factors influencing the pursuit of a criterion based, progressive rehabilitation program after distal biceps tendon repair. This commentary seeks to provide an update on current treatment strategies used in distal biceps rehabilitation with accompanying scientific rationale. Level of evidence 5.

Acromioclavicular (AC) joint injuries are a common cause of shoulder pain, particularly among young athletes participating in contact sports. Injuries to the AC joint most commonly occur from direct impact at the acromion and are classified as types I to VI. Although most AC joint injuries can be treated nonoperatively, types IV to VI are best treated with surgery, with type III being controversial and most surgeons recommending an initial trial of nonoperative treatment. Although numerous surgical techniques have been described, no gold standard technique has been established. Biomechanical testing suggests that anatomic reconstruction of both the AC and coracoclavicular ligaments results in a superior surgical construct. The objective of this Technical Note is to describe our preferred technique for the primary treatment of AC joint instability in the acute and chronic setting. Using 2 free tendon grafts in combination with a cortical button suspensory device combines the advantages of a nonrigid biologic and anatomic AC and coracoclavicular ligament reconstruction while benefiting from the strength of a cortical suspensory device in resisting displacement of the AC joint. © 2018 Arthroscopy Association of North America

Purpose To determine 2-year functional outcomes using an all-inside quadrupled semitendinosus anterior cruciate ligament (ACL) reconstruction technique in male and female patients. Methods A total of 100 patients who underwent quadrupled semitendinosus all-inside hamstring ACL reconstruction by a single surgeon were enrolled. Patient-reported outcome scores collected preoperatively and postoperatively included visual analog scale, Veterans Rand 12-Item Health Survey (VR-12; Physical and Mental), Marx Activity Scale, Knee Injury and Osteoarthritis Outcome Scores (KOOS), and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Results A total of 100 ACL reconstructions comprising 62 female, and 38 male patients were included in this study. Mean graft diameter was 9.4 mm in female and 9.8 mm in male patients (range, 9-11). Outcome scores demonstrated improvement from preoperative to 2-year postoperative respectively: visual analog scale pain 3.18, 1.07, VR-12 physical 36.35, 52.64, VR-12 mental 53.96, 54.65, KOOS pain 59.17, 89.03, KOOS symptoms 52.64, 80.79, KOOS Activities of Daily Living 69.38, 95.4, KOOS Sport 28.97, 81.25, KOOS Quality of Life 27.54, 71.56, WOMAC Pain 71.56, 92.65, WOMAC Stiffness 60.55, 84.13, and WOMAC Function 69.38, 95.4. Marx activity score decreased from baseline (10.98) to 2 years’ postoperatively (8.75). At 2 years, patient expectations were met or exceeded with regards to pain (94%), motion and strength (91%), normal function of daily living (95%), and return to sport (79%). Conclusions Anatomic all-inside quadrupled semitendinosus ACL reconstruction improves functional outcomes similarly to previous studies between baseline and clinical follow-up at 1-year and 2-years postoperatively and is comparable in both male and female patients. Level of Evidence Level III, retrospective comparative study.

Treatment of end-stage glenohumeral arthritis in young patients is a challenge; however, there is a lack of consensus on optimal treatment algorithms. A thorough history and physical examination are essential. Nonoperative treatments should first be attempted, whereas surgical options range from arthroscopic debridement to arthroplasty. One arthroplasty option is glenohumeral resurfacing with the objective of maintaining more native anatomy and bone stock. The described treatment includes a hemi-cap implant for the humerus and inlay polyethylene glenoid. While hemi-caps have been successfully used for decades, inlay glenoid implants are a more modern treatment, with the objective of less glenoid loosening, the typical complication and failure method in young patients. With the potential for greater longevity and preservation of anatomy, glenohumeral resurfacing for end-stage shoulder arthritis is an important treatment option to consider before total shoulder arthroplasty. This Technical Note describes resurfacing of the glenohumeral joint in a young, active patient presenting with extensive osteoarthritis on both the glenoid and humerus after a previous failed Trillat stabilization.

Nitrogen-containing bisphosphonates (N-BPs), such as alendronate, are the most widely prescribed medications for diseases involving bone, with nearly 200 million prescriptions written annually. Recently, widespread use of N-BPs has been challenged due to the risk of rare but traumatic side effects such as atypical femoral fracture (AFFs) and osteonecrosis of the jaw (ONJ). N-BPs bind to and inhibit farnesyl diphosphate synthase (FDPS), resulting in defects in protein prenylation. Yet it remains poorly understood what other cellular factors might allow N-BPs to exert their pharmacological effects. Here, we performed genome-wide studies in cells and patients to identify the poorly characterized gene, ATRAID. Loss of ATRAID function results in selective resistance to N-BP-mediated loss of cell viability and the prevention of alendronate-mediated inhibition of prenylation. ATRAID is required for alendronate inhibition of osteoclast function, and ATRAID-deficient mice have impaired therapeutic responses to alendronate in both postmenopausal and senile (old age) osteoporosis models. Lastly, we performed exome sequencing on patients taking N-BPs that suffered ONJ or an AFF. ATRAID is one of three genes that contain rare non-synonymous coding variants in patients with ONJ or AFF that is also differentially expressed in poor outcome groups of patients treated with N-BPs. We functionally validated this patient variation in ATRAID as conferring cellular hypersensitivity to N-BPs. Our work adds key insight into the mechanistic action of N-BPs and the processes that might underlie differential responsiveness to N-BPs in people.One Sentence SummaryATRAID is essential for responses to the commonly prescribed osteoporosis drugs nitrogen-containing bisphosphonates.OverlineBONE

ObjectivesRisk factors for anterior shoulder dislocation include young age, contact activities and male sex. The influence of sex on patient-reported outcomes of arthroscopic Bankart repair (ABR) is unclear, with few studies reporting potential differences. This study’s purpose was to compare patient-reported outcomes of males and females following ABR.MethodsProspectively collected data was analysed for 281 patients (males: 206, females: 75) after ABR with preoperative, 1-year and 2-year follow-up responses. The Wilcoxon signed-rank and χ2 tests, preoperative, 1 year and 2 year follow-up results were examined to determine differences of scores in males versus females.ResultsNo statistically significant sex differences were observed in Simple Shoulder Test (SST), American Shoulder and Elbow Surgeons (ASES), Visual Analogue Scale (VAS) or Single Assessment Numerical Evaluation (SANE) Scores at 1-year or 2-year follow-up. Females had lower Veterans RAND 12-item health survey (VR-12) mental health subscores at 2-year follow-up (females: 52.3±9.0, males: 55.8±7.6, p=0.0016). Females were more likely to report that treatment had ‘exceeded expectations’ at 2-year follow-up regarding motion, strength, function and normal sports activities.ConclusionResults of study demonstrate that ABR has similar outcomes for both males and females. There were no statistically significant sex-related differences in SST, ASES, VAS or SANE scores following ABR. VR-12 mental health subscores showed a minimal difference at 2-year follow-up, with lower scores in females.Level of evidenceRetrospective cohort study; level II.

The multiplicity of new therapies for breast cancer presents a challenge for treatment selection. We describe a 17-gene digital signature of breast circulating tumor cell (CTC)–derived transcripts enriched from blood, enabling high-sensitivity early monitoring of response. In a prospective cohort of localized breast cancer, an elevated CTC score after three cycles of neoadjuvant therapy is associated with residual disease at surgery (P = 0.047). In a second prospective cohort with metastatic breast cancer, baseline CTC score correlates with overall survival (P = 0.02), as does persistent CTC signal after 4 weeks of treatment (P = 0.01). In the subset with estrogen receptor (ER)–positive disease, failure to suppress ER signaling within CTCs after 3 weeks of endocrine therapy predicts early progression (P = 0.008). Drug-refractory ER signaling within CTCs overlaps partially with presence of ESR1 mutations, pointing to diverse mechanisms of acquired endocrine drug resistance. Thus, CTC-derived digital RNA signatures enable noninvasive pharmacodynamic measurements to inform therapy in breast cancer. Significance: Digital analysis of RNA from CTCs interrogates treatment responses of both localized and metastatic breast cancer. Quantifying CTC-derived ER signaling during treatment identifies patients failing to respond to ER suppression despite having functional ESR1. Thus, noninvasive scoring of CTC-RNA signatures may help guide therapeutic choices in localized and advanced breast cancer. Cancer Discov; 8(10); 1286–99. ©2018 AACR. This article is highlighted in the In This Issue feature, p. 1195

Blood-based biomarkers are critical in metastatic prostate cancer, where characteristic bone metastases are not readily sampled, and they may enable risk stratification in localized disease. We established a sensitive and high-throughput strategy for analyzing prostate circulating tumor cells (CTC) using microfluidic cell enrichment followed by digital quantitation of prostate-derived transcripts. In a prospective study of 27 patients with metastatic castration-resistant prostate cancer treated with first-line abiraterone, pretreatment elevation of the digital CTCM score identifies a high-risk population with poor overall survival (HR = 6.0; P = 0.01) and short radiographic progression-free survival (HR = 3.2; P = 0.046). Expression of HOXB13 in CTCs identifies 6 of 6 patients with ≤12-month survival, with a subset also expressing the ARV7 splice variant. In a second cohort of 34 men with localized prostate cancer, an elevated preoperative CTCL score predicts microscopic dissemination to seminal vesicles and/or lymph nodes (P < 0.001). Thus, digital quantitation of CTC-specific transcripts enables noninvasive monitoring that may guide treatment selection in both metastatic and localized prostate cancer. Significance: There is an unmet need for biomarkers to guide prostate cancer therapies, for curative treatment of localized cancer and for application of molecularly targeted agents in metastatic disease. Digital quantitation of prostate CTC-derived transcripts in blood specimens is predictive of abiraterone response in metastatic cancer and of early dissemination in localized cancer. Cancer Discov; 8(3); 288–303. ©2018 AACR. See related commentary by Heitzer and Speicher, p. 269. This article is highlighted in the In This Issue feature, p. 253

Significance Identifying predictive biomarkers of therapeutic response for melanoma patients treated with immune checkpoint inhibitors is a major challenge. By combining microfluidic enrichment for melanoma circulating tumor cells (CTCs) together with RNA-based droplet digital PCR quantitation, we have established a highly sensitive and robust platform for noninvasive, blood-based monitoring of tumor burden. Serial monitoring of melanoma patients treated with immune checkpoint inhibitors shows rapid changes in CTC score, which precede standard clinical assessment and are highly predictive of long-term clinical outcome. Early on-treatment digital monitoring of CTC dynamics may thus help identify patients likely to benefit from immune checkpoint inhibition therapy., A subset of patients with metastatic melanoma have sustained remissions following treatment with immune checkpoint inhibitors. However, analyses of pretreatment tumor biopsies for markers predictive of response, including PD-1 ligand (PD-L1) expression and mutational burden, are insufficiently precise to guide treatment selection, and clinical radiographic evidence of response on therapy may be delayed, leading to some patients receiving potentially ineffective but toxic therapy. Here, we developed a molecular signature of melanoma circulating tumor cells (CTCs) to quantify early tumor response using blood-based monitoring. A quantitative 19-gene digital RNA signature (CTC score) applied to microfluidically enriched CTCs robustly distinguishes melanoma cells, within a background of blood cells in reconstituted and in patient-derived (n = 42) blood specimens. In a prospective cohort of 49 patients treated with immune checkpoint inhibitors, a decrease in CTC score within 7 weeks of therapy correlates with marked improvement in progression-free survival [hazard ratio (HR), 0.17; P = 0.008] and overall survival (HR, 0.12; P = 0.04). Thus, digital quantitation of melanoma CTC-derived transcripts enables serial noninvasive monitoring of tumor burden, supporting the rational application of immune checkpoint inhibition therapies.

Purpose To determine whether single-strand semitendinosus autograft or allograft dimensions can reliably predict quadrupled graft diameter and length. Methods Intraoperative semitendinosus graft measurements were recorded for consecutive all-inside anterior cruciate ligament (ACL) reconstructions from 2013 to 2016 and retrospectively reviewed. Intraoperative single-strand tendon length and width and the corresponding quadrupled graft length and diameter were recorded. Pearson correlation coefficients were used to assess the linear association between single-strand width and quadrupled diameter, as well as between single-strand length and quadrupled length. Linear regression models were used to predict quadrupled values. To test the accuracy of the predicted quadrupled values, dimensions from an additional series of 30 all-inside ACL reconstructions were reviewed. Results Seventy-three ACL reconstruction procedures were reviewed. We excluded 12 grafts because gracilis and semitendinosus constructs were used. Thus 61 semitendinosus quadrupled grafts (30 autografts and 31 allografts) were included. Single-strand width was associated with quadrupled diameter ( P = .012), and single-strand length was associated with quadrupled graft length ( P Conclusions Quadrupled hamstring graft length and diameter may be accurately predicted based on length and width of the semitendinosus tendon used for all-inside, single-bundle ACL reconstruction. The ability to predict quadrupled graft dimensions can guide the surgeon in intraoperative decision making and ensure the desired ACL graft dimensions are achieved, thereby minimizing the risk of ACL reconstruction failure. Level of Evidence Level III, retrospective comparative study.

Introduction Non-small-cell lung cancer (NSCLC) is among the most common cancers, accounting for 85% of all lung cancer cases. Drug combinations are known to more effective than single drugs and can delay drug resistance. NSCLC has a poor prognosis and there are no approved targeted cancer drug combinations. Here we have conducted the first large-scale screen of drug combinations in NSCLC cell lines to identify new candidate synergistic combinations and map the landscape of synthetic lethal and synergistic pathways underlying them. Results We conducted the largest of its kind in vitro drug combination screening of 81 NSCLC cell-lines for 21 x 242 cancer drug combinations at 5 different doses. Our collection of approved and investigational drugs included mainly targeted drugs as well as some chemotherapy drugs. For each drug combination, we computed the synergy score using the Bliss model. Our key findings are as follows: 1. We identify drug combinations (like CHEK and WEE1 inhibitors) that are highly synergistic in many NSCLC cell lines. 2. Combinations of targeted therapies are overall more synergistic than chemotherapeutic drug combinations. 3. Synergistic combinations occur more within pathways. 4. We see that most drug combinations do not make cell lines more sensitive than the most sensitive cell line to the corresponding single agents across all the cell lines tested. 5. We identify a new class of drug combinations - called super-sensitizers - whose combined effects are larger than the effects observed for the individual drugs composing across all of the cohort cell-lines. 6. Combinations of multi-target drugs tend to be more synergistic than single-target drugs. 7. We identify and characterize a set of context-specific drug combinations that are differentially synergistic or differentially effective (i.e. viability after treatment) between wild-type vs mutant cell lines of various cancer drivers (KRAS, TP53, STK11, PIK3CA, EGFR). 8. Studying potential mechanistic underpinnings of drug synergism in NSCLC, we find that: (a) synergistic combinations target genes that have more proximal physical interactions, (b) some synergistic combinations target synthetic lethal combinations, (c) a subset of synergistic combinations have further support by NSCLC patients survival analysis. Conclusions We performed the first large in vitro drug combination screen in NSCLC. We identify top drug combinations that are highly synergistic and effective in many cell lines and characterize key biological properties and mechanisms underlying them. The screen results are provided in a user-friendly public data resource that can help facilitate the development of targeted drug combinations in lung cancer therapy. Citation Format: Nishanth Ulhas Nair, Adam Friedman, Patricia Greninger, Avinash D. Sahu, Ellen Murchie, Giovanna Stein Crowther, Joe McClanaghan, Joo Sang Lee, Daniel A. Haber, Jeffrey Engelman, Eytan Ruppin, Cyril Benes. A large cellular screen charting the landscape of synergistic drug combinations in lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1053.

Early detection strategies for pancreatic ductal adenocarcinoma are needed. This article describes a high‐sensitivity platform for detection of epithelial cells shed from preneoplastic lesions at a high risk of becoming malignant and a combination of technologies that could be used for early detection of disease in high‐risk patients., Background. Recent work has demonstrated early shedding of circulating epithelial cells (CECs) from premalignant intraductal papillary mucinous neoplasms (IPMNs). However, the potential use of CECs as a “liquid biopsy” for patients with IPMNs has been limited by antigen dependence of CEC isolation devices and the lack of robust detection biomarkers across CEC phenotypes. Materials and Methods. We utilized a negative depletion microfluidic platform to purify CECs from contaminating leukocytes and coupled this platform with immunofluorescence, RNA in situ hybridization, and RNA sequencing (RNA‐seq) detection and enumeration. Results. Using established protein (EpCAM, cytokeratins) and novel noncoding RNA (HSATII, cytokeratins) biomarkers, we detected CECs in 88% of patients bearing IPMN lesions. RNA‐seq analysis for MUC genes confirm the likely origin of these CECs from pancreatic lesions. Conclusion. Our findings increase the sensitivity of detection of these cells and therefore could have clinical implications for cancer risk stratification. Implications for Practice. This work describes a high‐sensitivity platform for detection of epithelial cells shed from preneoplastic lesions at high risk of malignant transformation. Further research efforts are underway to define the transcriptional programs that might allow discrimination between circulating cells released from tumors that will become malignant and cells released from tumors that will not. After further refinement, this combination of technologies could be deployed for monitoring and early detection of patients at high risk for developing new or recurrent pancreatic malignancies.

Context: Taping is commonly used in the management of several musculoskeletal conditions, including patellofemoral pain syndrome (PFPS). Specific guidelines for taping are unknown. Objective: To investigate the efficacy of knee taping in the management of PFPS. Our hypothesis was that tension taping and exercise would be superior to placebo taping and exercise as well as to exercise or taping alone. Data Sources: The PubMed/MEDLINE, Cochrane, Rehabilitation and Sports Medicine Source, and CINAHL databases were reviewed for English-language randomized controlled trials (RCTs) evaluating the efficacy of various taping techniques that were published between 1995 and April 2015. Keywords utilized included taping, McConnell, kinesio-taping, kinesiotaping, patellofemoral pain, and knee. Study Selection: Studies included consisted of RCTs (level 1 or 2) with participants of all ages who had anterior knee or patellofemoral pain symptoms and had received nonsurgical management using any taping technique. Study Design: Systematic review. Level of Evidence: Level 2. Data Extraction: A checklist method was used to determine selection, performance, detection, and attrition bias for each article. A quality of evidence grading was then referenced using the validated PEDro database for RCTs. Three difference comparison groups were compared: tension taping and exercise versus placebo taping and exercise (group 1), placebo taping and exercise versus exercise alone (group 2), and tension taping and exercise versus taping alone (group 3). Results: Five RCTs with 235 total patients with multiple intervention arms were included. Taping strategies included McConnell and Kinesiotaping. Visual analog scale (VAS) scores indicated improvement in all 3 comparison groups (group 1: 91 patients, 39% of total, mean VAS improvement 44.9 [tension taping + exercise] vs 66 [placebo taping + exercise]; group 2: 56 patients, 24% of total, mean VAS improvement 66 [placebo taping + exercise] vs 47.6 [exercise alone]; and group 3: 112 patients, 48% of total, mean VAS improvement 44.9 [tension taping + exercise] vs 14.1 [taping alone]). Conclusion: This systematic review supports knee taping only as an adjunct to traditional exercise therapy for PFPS; however, it does not support taping in isolation.

Background: Metastatic breast cancer (MBC) accounts for most breast cancer deaths. In 2018, two Poly(ADP-ribose) polymerase (PARP) inhibitors, olaparib and talazoparib, were approved for the treatment of MBC patients with germline BRCA1 or 2 mutations based on the OlympiAD and EMBRACA trials (Robson, NEJM, 2017 and Litton, NEJM, 2018), which demonstrated improvement in progression-free survival (PFS) with PARP inhibitors compared to chemotherapy. These landmark findings have triggered much interest in studying PARP inhibitors in MBC, but germline BRCA mutations only account for 5-10% of breast cancer patients. A critical question is whether PARP inhibitors may also be beneficial in MBC with somatic BRCA1 or 2 mutations, similar to somatic BRCA mutant ovarian cancer where they are effective(George, Oncotarget, 2017). Liquid biopsies via circulating cell-free DNA (cfDNA) may unveil somatic mutations. We previously demonstrated (Vidula, SABCS, PD1-13, 2017) that a subset of MBC patients have detectable somatic cfDNA BRCA1 or 2 mutations, in the absence of germline BRCA mutations, highlighting the role of liquid biopsies for detection of somatic BRCA mutations in MBC. Based on our preliminary data, we hypothesize that we can utilize liquid biopsies to detect somatic BRCA1 or 2 mutations and guide therapy selection with a PARP inhibitor in patients with MBC. Methods: In this single arm phase II clinical trial we will evaluate the efficacy and safety of a PARP inhibitor, talazoparib, in patients with somatic BRCA mutant MBC (N=30). Patients will undergo cfDNA screening to identify cfDNA somatic BRCA1 or 2 deleterious mutations. Patients without known BRCA germline mutations who have cfDNA BRCA1 or 2 mutations will be enrolled at the Massachusetts General Hospital and University of California San Francisco. The study will enroll both patients with hormone receptor positive/HER2 negative breast cancer and triple-negative breast cancer whose disease has progressed on at least 1 prior line of endocrine therapy and at least 1 prior line of chemotherapy for MBC, respectively. Patients may have received any number of prior lines of therapy. Eligible patients will be treated with talazoparib 1 mg daily until progression (evaluated with serial CT chest/abdomen/pelvis and bone scan) or unacceptable toxicity. The primary endpoint is to determine PFS by RECIST 1.1. Patients will be enrolled in a two-stage design, providing 80% power to demonstrate that the treatment is associated with “success” (PFS > 12 weeks) in ≥53% patients (4% alpha). Secondary endpoints include objective response rate and safety/tolerability (NCI CTCAE v5.0). Serial cfDNA will be collected at baseline and during treatment to evaluate for changes in the BRCA cfDNA mutant allelic fraction in response to therapy. The impact of pre-existing resistance mutations in baseline cfDNA, particularly BRCA reversion mutations that can occur with prior platinum exposure (Weigelt, CCR, 2017), on outcomes will be studied. Paired liquid biopsies (pre- and post-treatment) will be compared to generate hypotheses about potential novel targets for future combination studies with talazoparib to improve response(exploratory objective). This study may help expand the applicability of talazoparib in MBC. This study was activated in July 2019. Funding support for the clinical trial is provided by a Pfizer ASPIRE award. (NCT03990896). Citation Format: Neelima Vidula, Nora Horick, Erin Basile, Sara Sutherland, Ruth Fax, Daniel Haber, Leif Ellisen, Hope S. Rugo, Aditya Bardia. Evaluation of talazoparib, a PARP inhibitor, in patients with somatic BRCA mutant metastatic breast cancer: Genotyping based clinical trial [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr OT2-03-03.

Objectives: The instability severity index score (ISIS) was designed to predict the risk of recurrence after arthroscopic instability shoulder surgery and to better predict those who would benefit from an open or bone transfer operation. Although this score has been widely disseminated to predict recurrence, there are certain areas in which preoperative assessment is limited, especially in radiographic workup. The objective of this study was to examine the validity of ISIS based on its existing variables, as well as to evaluate additional imaging and patient history variables pertinent to the potential redevelopment of a new score to assess risk of recurrent anterior instability following an arthroscopic Bankart repair. Methods: All consecutive patients were prospectively enrolled with recurrent anterior shoulder instability who subsequently underwent an arthroscopic stabilization with minimum 24 months follow-up. Exclusion criteria included, prior surgery on the shoulder, posterior or multidirectional instability, or a rotator cuff tear. All instability severity index score variables were recorded (age Results: There were 217 consecutive patients (209 male-96.5%, 8 female-3.5%) who met criteria and were all treated with a primary arthroscopic shoulder stabilization during a 3.5-year period (2007-2011), with mean follow-up of 42 (range, 26-58 mos). The mean age at first instability event was 23.9 (range, 16-48), with 55% right shoulder affected, 60% dominant shoulder. Outcomes were improved from mean scores preoperative (WOSI=1050/2100, ASES=61.0, SANE=52.5) to postoperative (WOSI=305/2100, ASES=93.5, SANE=95.5), and 11.5% (25/217) had evidence or recurrent instability or subluxation. A total of 51/217 were 20 years or under, hyperlaxity in 5, Hill Sachs on internal rotation XR in 77, glenoid contour on AP XR in 41, with an overall mean ISIS score of 3.6. Factors associated with failure were glenoid bone loss greater than 14.5%(p11.5 months(p 1.3 cm3 with H>1.5 cm, W>1.0 cm and D>5 mm(p0.45), sports participation, and Instability Severity Score (mean=3.4 success, vs 3.9 failure, p>0.44). Conclusion: At nearly four years of follow-up, there was an 11.5% failure rate of scope stabilization surgery. However, there was no correlation between treatment outcome and the ISIS measure given a mean score of 3.4 with little difference identified in those that failed. However, several important parameters previously unidentified were detected including, glenoid bone loss >14.5%, Hill Sachs volume >1.3 cm3, and time length of instability symptoms. Therefore, the ISIS measure may need to be redesigned in order to incorporate variables that more accurately portray the actual risk of failure following arthroscopic stabilization.

Articular cartilage injuries of the knee are common among young, active patients presenting with knee pain, swelling, and/or mechanical symptoms. These injuries have limited healing potential due to the avascular nature of hyaline cartilage. While several treatment options exist, osteochondral allograft (OCA) transplantation for the knee has been used successfully in primary management of large chondral or osteochondral defects and salvage of previously failed cartilage repair. OCA transplantation potentially yields a natural, matching contour of the native recipient surface anatomy and transplants mature, viable hyaline cartilage to the affected defect. Following OCA transplantation, strict compliance with a rehabilitation protocol is essential to enable optimal recovery. The outlined rehabilitation protocol is informed by the existing literature and incorporates current rehabilitation principles, the science of osteochondral incorporation, and adaptations based on an individual's readiness to progress through subsequent phases. The purpose of this clinical commentary is to discuss the diagnosis, surgical management, and post-operative rehabilitation following OCA transplantation and to assist the physical therapist in returning athletes to full sports participation.

Solid tumors release a variety of cellular components into the bloodstream that can be sampled by liquid biopsy. These blood-borne biomarkers include circulating tumor cells, exosomes, cell-free DNA, and cell-free noncoding RNA. As hepatocellular carcinoma (HCC) grows in worldwide prevalence and mortality, the application of emerging liquid biopsy technologies to HCC has been a rapidly advancing area of research. The qualitative and quantitative analysis of these biomarkers holds promise for improving the clinical care of individuals with HCC. In particular, research has focused on the role of liquid biopsy in the detection of HCC at an early and curable stage, given the major clinical need for biomarkers for this purpose. Panels of liquid biopsy biomarkers or combinations of conventional and novel assays will likely optimize sensitivity and specificity in HCC surveillance for at-risk populations. Additional applications of liquid biopsy in HCC include assessment of prognosis, monitoring treatment response and disease recurrence, and assaying underlying tumor biology. The future of liquid biopsy research will move the field from exploratory studies with heterogenous techniques to validated high-throughput molecular analyses and eventually from bench to bedside.

Circulating tumour cells in women with advanced oestrogen-receptor (ER)-positive/human epidermal growth factor receptor 2 (HER2)-negative breast cancer acquire a HER2-positive subpopulation after multiple courses of therapy. In contrast to HER2-amplified primary breast cancer, which is highly sensitive to HER2-targeted therapy, the clinical significance of acquired HER2 heterogeneity during the evolution of metastatic breast cancer is unknown. Here we analyse circulating tumour cells from 19 women with ER+/HER2- primary tumours, 84% of whom had acquired circulating tumour cells expressing HER2. Cultured circulating tumour cells maintain discrete HER2+ and HER2- subpopulations: HER2+ circulating tumour cells are more proliferative but not addicted to HER2, consistent with activation of multiple signalling pathways; HER2- circulating tumour cells show activation of Notch and DNA damage pathways, exhibiting resistance to cytotoxic chemotherapy, but sensitivity to Notch inhibition. HER2+ and HER2- circulating tumour cells interconvert spontaneously, with cells of one phenotype producing daughters of the opposite within four cell doublings. Although HER2+ and HER2- circulating tumour cells have comparable tumour initiating potential, differential proliferation favours the HER2+ state, while oxidative stress or cytotoxic chemotherapy enhances transition to the HER2- phenotype. Simultaneous treatment with paclitaxel and Notch inhibitors achieves sustained suppression of tumorigenesis in orthotopic circulating tumour cell-derived tumour models. Together, these results point to distinct yet interconverting phenotypes within patient-derived circulating tumour cells, contributing to progression of breast cancer and acquisition of drug resistance.

Purpose: The T790M gatekeeper mutation in the EGFR is acquired by some EGFR-mutant non–small cell lung cancers (NSCLC) as they become resistant to selective tyrosine kinase inhibitors (TKI). As third-generation EGFR TKIs that overcome T790M-associated resistance become available, noninvasive approaches to T790M detection will become critical to guide management. Experimental Design: As part of a multi-institutional Stand-Up-To-Cancer collaboration, we performed an exploratory analysis of 40 patients with EGFR-mutant tumors progressing on EGFR TKI therapy. We compared the T790M genotype from tumor biopsies with analysis of simultaneously collected circulating tumor cells (CTC) and circulating tumor DNA (ctDNA). Results: T790M genotypes were successfully obtained in 30 (75%) tumor biopsies, 28 (70%) CTC samples, and 32 (80%) ctDNA samples. The resistance-associated mutation was detected in 47% to 50% of patients using each of the genotyping assays, with concordance among them ranging from 57% to 74%. Although CTC- and ctDNA-based genotyping were each unsuccessful in 20% to 30% of cases, the two assays together enabled genotyping in all patients with an available blood sample, and they identified the T790M mutation in 14 (35%) patients in whom the concurrent biopsy was negative or indeterminate. Conclusions: Discordant genotypes between tumor biopsy and blood-based analyses may result from technological differences, as well as sampling different tumor cell populations. The use of complementary approaches may provide the most complete assessment of each patient's cancer, which should be validated in predicting response to T790M-targeted inhibitors. Clin Cancer Res; 22(5); 1103–10. ©2015 AACR.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most devastating and painful cancers. It is often highly resistant to therapy owing to inherent chemoresistance and the desmoplastic response that creates a barrier of fibrous tissue preventing transport of chemotherapeutics into the tumor. The growth of the tumor in pancreatic cancer often leads to invasion of other organs and partial or complete biliary obstruction, inducing intense pain for patients and necessitating tumor resection or repeated stenting. Here, we have developed a delivery device to provide enhanced palliative therapy for pancreatic cancer patients by providing high concentrations of chemotherapeutic compounds locally at the tumor site. This treatment could reduce the need for repeated procedures in advanced PDAC patients to debulk the tumor mass or stent the obstructed bile duct. To facilitate clinical translation, we created the device out of currently approved materials and drugs. We engineered an implantable poly(lactic-co-glycolic)-based biodegradable device that is able to linearly release high doses of chemotherapeutic drugs for up to 60 days. We created five patient-derived PDAC cell lines and tested their sensitivity to approved chemotherapeutic compounds. These in vitro experiments showed that paclitaxel was the most effective single agent across all cell lines. We compared the efficacy of systemic and local paclitaxel therapy on the patient-derived cell lines in an orthotopic xenograft model in mice (PDX). In this model, we found up to a 12-fold increase in suppression of tumor growth by local therapy in comparison to systemic administration and reduce retention into off-target organs. Herein, we highlight the efficacy of a local therapeutic approach to overcome PDAC chemoresistance and reduce the need for repeated interventions and biliary obstruction by preventing local tumor growth. Our results underscore the urgent need for an implantable drug-eluting platform to deliver cytotoxic agents directly within the tumor mass as a novel therapeutic strategy for patients with pancreatic cancer. Keywords: Pancreatic cancer; Chemoresistance; Local delivery; Patient-derived xenograft; Paclitaxel; Poly(lactic-co-glycolic acid), National Institutes of Health (U.S.) (Grant P30-CA14051)

Women with metastatic hormone receptor positive (HR+) breast cancer benefit from therapies targeting the Estrogen Receptor (ER), but the tumors eventually acquire resistance leading to disease progression. Endocrine resistance involves multiple mechanisms, including deregulation of ER signaling due to activating mutations in the ligand-binding domain of the ESR1 gene. In addition, recent reports describe specific contexts of ESR1 mutations in the preclinical setting where tamoxifen and fulvestrant have demonstrated limited activity. These data together with the frequency of ESR1 mutations (~20-60%) observed in mBC patients in the clinic suggest a need for more effective agents. The activity of elacestrant (RAD1901), an oral selective estrogen receptor degrader (SERD) with demonstrated activity against tumors harboring ESR1 mutations, both in preclinical models and in patients with previously treated advanced breast cancer, was assessed in ex vivo cultures of circulating tumor cells (CTCs) derived from women with metastatic HR+ breast cancer who had received multiple prior hormone therapies. Cultured breast CTCs underwent apoptosis following exposure to elacestrant, and CTCs harboring mutant ESR1 were significantly growth inhibited (IC50: 17.65 ± 3.32 nM) by elacestrant compared to the standard-of-care (SOC) agent fulvestrant. Furthermore, elacestrant eliminated the fulvestrant-resistant CTC populations in vitro. Elacestrant treatment led to significant suppression of ER target genes, PGR and GREB1. Elacestrant is currently being investigated versus standard of care endocrine monotherapy in a Phase 3 clinical trial (EMERALD) in patients with ER+/HER2- advanced/metastatic breast cancer. A co-primary endpoint of the EMERALD trial is evaluation of progression-free survival in patients whose tumors harbor ESR1 mutations. Citation Format: Taronish Dorab Dubash, Aditya Bardia, Brittany A Reeves, Brian Chirn, Annamaria Szabolcs, Ben S Wittner, John Iafrate, David Ting, Hitisha K Patel, Teeru Bihani, Mehmet Toner, Daniel A Haber, Shyamala Maheswaran. Elacestrant (RAD1901) inhibits growth of ex vivo cultured circulating tumor cells derived from hormone receptor-positive metastatic breast cancer (mBC) patients including those harboring ESR1 mutations [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P4-01-06.

Significance PDAC epithelial (E) and quasi-mesenchymal (QM) subtypes can be modulated by different therapies demonstrating the plasticity of PDAC cells that contributes to the heterogeneity of PDAC tumors and their intrinsic resistance to a broad spectrum of therapies. Understanding and monitoring the fluidity of PDAC E and QM states are critical to the development of improved clinical trial design to target these different subpopulations., Transcriptional profiling has defined pancreatic ductal adenocarcinoma (PDAC) into distinct subtypes with the majority being classical epithelial (E) or quasi-mesenchymal (QM). Despite clear differences in clinical behavior, growing evidence indicates these subtypes exist on a continuum with features of both subtypes present and suggestive of interconverting cell states. Here, we investigated the impact of different therapies being evaluated in PDAC on the phenotypic spectrum of the E/QM state. We demonstrate using RNA-sequencing and RNA-in situ hybridization (RNA-ISH) that FOLFIRINOX combination chemotherapy induces a common shift of both E and QM PDAC toward a more QM state in cell lines and patient tumors. In contrast, Vitamin D, another drug under clinical investigation in PDAC, induces distinct transcriptional responses in each PDAC subtype, with augmentation of the baseline E and QM state. Importantly, this translates to functional changes that increase metastatic propensity in QM PDAC, but decrease dissemination in E PDAC in vivo models. These data exemplify the importance of both the initial E/QM subtype and the plasticity of E/QM states in PDAC in influencing response to therapy, which highlights their relevance in guiding clinical trials.

Background Microfluidic platforms have demonstrated the ability to isolate rare circulating tumor cells from a wide variety of cancers. Our group has recently shown the ability to isolate circulating tumor cells (CTCs) from hepatocellular carcinoma (HCC) patients using a hematopoietic cell depletion microfluidic platform. However, the relationship of CTC generation and HCC progression is still not well understood. Tumor size is often used as a clinical prognostic factor, but there has been an inconsistent relationship of tumor size and metastatic recurrence. Characterizing the relationship of primary tumor size and CTCs would provide a better understanding of HCC tumor size and metastatic potential. Results CTCs in a cohort of HCC patients with quantitative tumor volume analysis was performed to determine if there was a relationship of tumor size to CTC generation. A total of 24 tumor volumetric analyses were used in this study, and a cutoff of 25 cc was used to create a low and high tumor volume group (median 5.56 vs 108 cc; p

Molecular drivers underlying bone metastases in human cancer are not well understood, in part due to constraints in bone tissue sampling. Here, RNA sequencing was performed of circulating tumor cells (CTC) isolated from blood samples of women with metastatic estrogen receptor (ER)+ breast cancer, comparing cases with progression in bone versus visceral organs. Among the activated cellular pathways in CTCs from bone-predominant breast cancer is androgen receptor (AR) signaling. AR gene expression is evident, as is its constitutively active splice variant AR-v7. AR expression within CTCs is correlated with the duration of treatment with aromatase inhibitors, suggesting that it contributes to acquired resistance to endocrine therapy. In an established breast cancer xenograft model, a bone-tropic derivative displays increased AR expression, whose genetic or pharmacologic suppression reduces metastases to bone but not to lungs. Together, these observations identify AR signaling in CTCs from women with bone-predominant ER+ breast cancer, and provide a rationale for testing androgen inhibitors in this subset of patients. Implications: This study highlights a role for the AR in breast cancer bone metastasis, and suggests that therapeutic targeting of the AR may benefit patients with metastatic breast cancer. Mol Cancer Res; 16(4); 720–7. ©2018 AACR.

Injuries to the acromioclavicular (AC) joint are common, particularly in the young and active population. Approximately 9% of all shoulder girdle injuries involve the AC joint, and AC joint dislocations represent approximately 8% of all joint dislocations throughout the body. AC joint injuries are graded as type I through type VI according to the Rockwood classification. Type I and II injuries are typically treated nonoperatively, whereas type IV, V, and VI injuries are most often treated surgically. A variety of surgical techniques have been described, including anatomic and nonanatomic reconstruction. However, up to 80% of patients go on to lose radiographic reduction, and between 20% and 30% have complications leading to reoperation. Therefore, the objective of this Technical Note is to describe our preferred technique for the treatment of AC joint instability in the revision setting. This technique uses a Dog Bone Button (Arthrex, Naples, FL) and 2 allografts.

Modeling the hematogenous spread of cancer cells to distant organs poses one of the greatest challenges in the study of human metastasis. Both tumor cell–intrinsic properties as well as interactions with reactive stromal cells contribute to this process, but identification of relevant stromal signals has been hampered by the lack of models allowing characterization of the metastatic niche. Here, we describe an implantable bioengineered scaffold, amenable to in vivo imaging, ex vivo manipulation, and serial transplantation for the continuous study of human metastasis in mice. Orthotopic or systemic inoculation of tagged human cancer cells into the mouse leads to the release of circulating tumor cells into the vasculature, which seed the scaffold, initiating a metastatic tumor focus. Mouse stromal cells can be readily recovered and profiled, revealing differential expression of cytokines, such as IL1β, from tumor-bearing versus unseeded scaffolds. Finally, this platform can be used to test the effect of drugs on suppressing initiation of metastatic lesions. This generalizable model to study cancer metastasis may thus identify key stromal-derived factors with important implications for basic and translational cancer research. Cancer Res; 74(24); 7229–38. ©2014 AACR.

To (1) describe the magnetic resonance imaging (MRI) characteristics of knee chondral injuries identified at the National Football League (NFL) Combine and (2) assess in-game performance of prospective NFL players with previously untreated knee chondral injuries and compare it with matched controls.All players with knee chondral injuries identified at the NFL Combine (2009-2015) were retrospectively reviewed. Players with prior knee surgery were excluded. A knee MRI for each player was reviewed; location, modified International Cartilage Repair Society (ICRS) grade (I-IV), and associated compartment subchondral edema were documented. Position, respective NFL Draft pick selection number, games started, played, snap percentage, and position-specific performance metrics during the first 2 NFL seasons were recorded for the injury and injury-free control group composed of players with (1) no prior knee injury, (2) no significant missed time prior to the NFL (≤2 total missed games in college), (3) no history of knee surgery, and (4) drafted in the respective NFL Draft following the NFL Combine.Of the 2,285 players reviewed, 101 (4.4%) had an injury without prior knee surgery. The patella (63.4%) and trochlea (34%) were most commonly affected. Defensive linemen were at highest risk for unrecognized injuries (odds ratio 1.8, P = .015). Players with previously untreated injuries, compared with controls, were picked later (mean pick: 125.8) and played (mean: 23) and started (mean: 10.4) fewer games during the initial 2 NFL seasons (P.001 for all). Particularly, subchondral bone edema and full-thickness cartilage injuries were associated with fewer games played (P = .003).The patellofemoral joint was most commonly affected in NFL Combine participants. Previously untreated knee articular injuries in players at the NFL Combine are associated with poorer early NFL performance in comparison to uninjured players. Subchondral bone edema and full-thickness cartilage injury on MRI were associated with fewer games played during the initial NFL career.Level III, case-control study.

With increasing shoulder instability events, the likelihood of a bony lesion of the glenoid and/or humeral head rises. Although bone loss of either the glenoid or humeral head may result in recurrent instability, bipolar lesions have been shown, in particular, to result in a negative and additive effect on glenohumeral stability. In the case of a bipolar lesion comprising severe glenoid bone loss and an engaging, "off-track" Hill-Sachs lesion, the bony foundation of the glenohumeral joint is compromised and bony augmentation is necessary. We present our preferred technique, made up of the application of a distal tibia allograft to address the glenoid bone loss and humeral head allograft to address the Hill-Sachs lesion, for the treatment of a severe bipolar lesion in the setting of recurrent anterior shoulder instability after a failed Latarjet procedure.

Circulating tumor cells (CTCs) are shed into the bloodstream by invasive cancers, but the difficulty inherent in identifying these rare cells by microscopy has precluded their routine use in monitoring or screening for cancer. We recently described a high-throughput microfluidic CTC-iChip, which efficiently depletes hematopoietic cells from blood specimens and enriches for CTCs with well-preserved RNA. Application of RNA-based digital PCR to detect CTC-derived signatures may thus enable highly accurate tissue lineage-based cancer detection in blood specimens. As proof of principle, we examined hepatocellular carcinoma (HCC), a cancer that is derived from liver cells bearing a unique gene expression profile. After identifying a digital signature of 10 liver-specific transcripts, we used a cross-validated logistic regression model to identify the presence of HCC-derived CTCs in nine of 16 (56%) untreated patients with HCC versus one of 31 (3%) patients with nonmalignant liver disease at risk for developing HCC (P < 0.0001). Positive CTC scores declined in treated patients: Nine of 32 (28%) patients receiving therapy and only one of 15 (7%) patients who had undergone curative-intent ablation, surgery, or liver transplantation were positive. RNA-based digital CTC scoring was not correlated with the standard HCC serum protein marker alpha fetoprotein (P = 0.57). Modeling the sequential use of these two orthogonal markers for liver cancer screening in patients with high-risk cirrhosis generates positive and negative predictive values of 80% and 86%, respectively. Thus, digital RNA quantitation constitutes a sensitive and specific CTC readout, enabling high-throughput clinical applications, such as noninvasive screening for HCC in populations where viral hepatitis and cirrhosis are prevalent.

Most cancers are detected when they cause symptoms that lead to medical evaluation. Unfortunately, in too many cases this results in diagnosis of cancers that are locally invasive or already metastatic and hence no longer curable with surgical resection or radiation treatment. Medical therapies, which might be curative in the setting of minimal tumor burden, typically provide more limited benefit in more advanced cancers, given the emergence of drug resistance ( 1 ). On page 926 of this issue, Cohen et al. ( 2 ) describe a strategy for early cancer detection, CancerSEEK, aimed at screening for multiple different cancers within the general population. This study challenges current assumptions in the field of blood-based biomarkers and sets the stage for the next generation of cancer screening initiatives.

Objectives: To report patient characteristics, fracture types, treatment methods, and clinical outcomes of children and adolescents treated for tibial tubercle fractures. Methods: Patients age 19 and under treated for tibial tubercle fractures at a single institution from 1995 to 2015 were identified and their medical records were retrospectively reviewed. Clinical and radiographic outcomes were reported for patients with a minimum follow up of 6 months. Associations were tested using Fisher’s exact test. Results: Two-hundred thirty seven tibial tubercle fractures were identified in 229 patients; 198 (86%) male. There were 139 (59%) left sided injuries. Mean age was 14.3 years (range 4.9 - 19.8). Mean body mass index (BMI) was 25. Osgood-Schlatters was identified in 72 (30%) cases and was most commonly associated with Ogden type I fractures (pConclusion: Tibial tubercle fractures occur most commonly in adolescent male athletes and are associated with basketball, elevated BMI, Osgood-Schlatters, and a risk of compartment syndrome. Following treatment, most patients return to sport. As the largest single series of tibial tubercle fractures reported, this study provides valuable insight into injury epidemiology, relationship with Osgood Schlatters, and clinical outcomes.

Epithelial cells in the circulation (circulating epithelial cells, or CECs) are analyzed as a non-invasive method to detect cancers; we investigated whether analysis of hepatocytes in the circulation can identify patients with chronic liver disease or hepatocellular carcinoma (HCC). We previously developed a cell-sorting device to isolate CECs from patient blood samples and combined it with an mRNA analysis system to identify CECs with liver-specific markers. We tested the ability of this device to detect CECs of hepatocyte origin in blood samples from healthy individuals (n=10), patients with chronic liver disease without HCC (n=39), and patients with HCC (n=54), using immunofluorescence. We confirmed hepatocyte origin using RNA-sequencing analysis. We found a similar concentration of circulating hepatocytes in blood samples from patients with chronic liver disease and HCC but an increased concentration from patients with advanced fibrosis compared to those without advanced fibrosis. Circulating hepatocytes isolated from patients with HCC had a different gene expression profile than those from patients with chronic liver disease. This system for detecting and analyzing circulating hepatocytes might be used in the evaluation of benign and malignant liver disease.

Background: Cabozantinib is an oral MET/VEGFR2 inhibitor. A recent phase II study of cabozantinib (100 mg daily) showed improved bone scans in subjects with metastatic castration-resistant prostate cancer (mCRPC), but adverse events (AE) caused frequent dose reductions. This study was designed to determine the efficacy and tolerability of cabozantinib at lower starting doses. Experimental Design: An adaptive design was used to determine the lowest active daily dose among 60, 40, and 20 mg. The primary endpoint was week 6 bone scan response, defined as ≥30% decrease in bone scan lesion area. The secondary endpoint was change in circulating tumor cells (CTC). Results: Among 11 evaluable subjects enrolled at 40 mg, there were 9 partial responses (PR), 1 complete response, and 1 stable disease (SD). Of 10 subjects subsequently enrolled at 20 mg, there were 1 PR, 5 SDs, and 4 with progressive disease. Among 13 subjects enrolled on the 40 mg expansion cohort, there were 6 PRs and 7 SDs. No subjects required dose reduction or treatment interruption at 6 or 12 weeks; 3 subjects at dose level 0 discontinued due to AEs by 12 weeks. At 40 mg, median treatment duration was 27 weeks. 58% of subjects with ≥5 CTCs/7.5mL at baseline converted to Conclusions: Cabozantinib 40 mg daily was associated with a high rate of bone scan response. Cabozantinib 40 mg daily was associated with better tolerability than previously reported for cabozantinib 100 mg daily. These observations informed the design of phase III studies of cabozantinib in mCRPC. Clin Cancer Res; 19(11); 3088–94. ©2013 AACR.

The shortage of healthcare workers caring for South Africa's 5-6 million persons living with HIV/AIDS (PLHA) calls for inquiry into workers' challenges and experiences. This exploratory study examines one little-studied challenge: stigmatisation of HIV/AIDS healthcare workers based on their association with PLHA. The authors tested the hypotheses that HIV/AIDS healthcare workers experience stigmatisation due to their association with PLHA, and that such association stigma is correlated with thoughts of leaving the HIV/AIDS field. A sample of 100 participants who provided direct care to PLHA was recruited from a variety of public and private HIV/AIDS care centres in Eastern Cape province, South Africa. Participants attended one of 12 focus groups held between June and August, 2008. They completed a 17-item questionnaire and discussed each item. Findings exhibit the presence of an adverse differentiation and labelling of HIV/AIDS healthcare workers, leading to status loss and discrimination, creating an impetus for HIV/AIDS healthcare workers to leave AIDS work altogether. A significant relationship (χ(2) (TREND) = 3.86, df = 1, P = 0.049) was found between contemplation of leaving AIDS work and perception of others' responses to their work with PLHA. In addition, associations emerged between type of AIDS worker and contemplation of working in AIDS care outside of South Africa (Kruskal-Wallis χ(2) = 6.96, df = 2, P = 0.031), with doctors and nurses reporting higher frequency of contemplating leaving South Africa to work with PLHA elsewhere (Mann-Whitney z = -2.53, P = 0.011). The study lays the foundation for additional research on the effects of association stigma. In turn, increased efforts to retain and recruit new HIV/AIDS healthcare workers will expand the pool of healthcare personnel to PLHA.

Background: Jones fractures result in subsequent dysfunction and remain an issue for athletes. Purpose: To (1) describe the epidemiology, treatment, and impact of Jones fractures identified at the National Football League (NFL) Scouting Combine on players’ early careers and (2) establish the value of computed tomography (CT) to determine bony healing after a fracture in prospective players. Study Design: Cohort study; Level of evidence, 3. Methods: All players who attended the combine between 2009 and 2015 were retrospectively reviewed to identify their history of Jones fractures. The playing position, treatment method, and number of missed collegiate games were recorded. The mean overall draft pick number, number of games started and played, snap percentage, and position-specific performance scores (fantasy score) over the first 2 years in the NFL were compared between players with fractures and controls. An imaging classification system was applied based on grading of each quadrant of the fifth metatarsal (plantar, dorsal, medial, lateral), with a score of 0 for not healed or 1 for healed. Results: Overall, the number of Jones fractures identified was 72 in 2285 athletes (3.2%), with all treated via intramedullary screw fixation. The mean overall draft pick number for players with fractures was 111.2 ± 67.9 compared with 99.0 ± 65.9 for controls ( P = .12). Performance scores for players with fractures were lower than those for controls across all positions, with a significant difference in running backs (2.6 vs 4.0, respectively; P < .001) and defensive linemen (1.4 vs 2.3, respectively; P = .02). The mean CT score was 2.5 ± 1.3. Of the 32 athletes who underwent imaging, 16 Jones fractures (50.0%) were healed or nearly healed, 12 (37.5%) were partially healed, and 4 (12.5%) showed little or no healing. The plantar cortex demonstrated the least healing (18/32; 56.3%), followed by the lateral cortex (15/32; 46.9%). Players with a mean score Conclusion: Based on CT, 50% of all players with a previous Jones fracture demonstrated incomplete healing. Moreover, position-specific performance scores over the first 2 years of a player’s career were lower across all positions for those with fractures compared with controls. Players with CT scores

A subset of patients with metastatic melanoma have sustained remissions following treatment with immune checkpoint inhibitors. However, analyses of pretreatment tumor biopsies for markers predictive of response, including PD-L1 expression and mutational burden, are insufficiently precise to guide treatment selection and clinical radiographic evidence of response on therapy may be delayed, leading to some patients receiving potentially ineffective but toxic therapy. Here, we developed a molecular signature of melanoma Circulating Tumor Cells (CTCs) to quantify early tumor response using blood-based monitoring. A quantitative 19-gene digital RNA signature (CTC-Score) applied to microfluidically-enriched CTCs robustly distinguishes melanoma cells, within a background of blood cells in reconstituted and in patient-derived (N=42) blood specimens. In a prospective cohort of 49 patients treated with immune checkpoint inhibitors, a decrease in CTC-Score within 7 weeks of therapy correlates with marked improvement in progression-free survival (Hazard Ratio (HR): 0.17, P=0.008) and overall survival (HR: 0.12, P=0.04). Thus, digital quantitation of melanoma CTC-derived transcripts enables serial noninvasive monitoring of tumor burden, supporting the rational application of immune checkpoint inhibition therapies. Citation Format: Xin Hong, Ryan J. Sullivan, Mark Kalinich, Tanya Kwan, Shiwei Pan, Joseph A. LiCausi, John D. Milner, Linda T. Nieman, Ben S. Wittner, Uyen Ho, Tianqi Chen, Ravi Kapur, Don Lawrence, Keith T. Flaherty, Lecia V. Sequist, Sridhar Ramaswamy, David T. Miyamoto, Michael Lawrence, Anita Giobbie-Hurder, Mehmet Toner, Kurt J. Isselbacher, Shyamala Maheswaran, Daniel A. Haber. Molecular signatures of circulating melanoma cells for monitoring early response to immune checkpoint therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-144.