Differential DNA damage repair and PARP inhibitor vulnerability of the mammary epithelial lineages.

It is widely assumed that all normal somatic cells can equally perform homologous recombination (HR) and non-homologous end joining in the DNA damage response (DDR). Here, we show that the DDR in normal mammary gland inherently depends on the epithelial cell lineage identity. Bioinformatics, post-irradiation DNA damage repair kinetics, and clonogenic assays demonstrated luminal lineage exhibiting a more pronounced DDR and HR repair compared to the basal lineage. Consequently, basal progenitors were far more sensitive to poly(ADP-ribose) polymerase inhibitors (PARPis) in both mouse and human mammary epithelium. Furthermore, PARPi sensitivity of murine and human breast cancer cell lines as well as patient-derived xenografts correlated with their molecular resemblance to the mammary progenitor lineages. Thus, mammary epithelial cells are intrinsically divergent in their DNA damage repair capacity and PARPi vulnerability, potentially influencing the clinical utility of this targeted therapy.

Mammary epithelial cells have lineage-rooted metabolic identities.

Cancer metabolism adapts the metabolic network of its tissue of origin. However, breast cancer is not a disease of a single origin. Multiple epithelial populations serve as the culprit cell of origin for specific breast cancer subtypes, yet our knowledge of the metabolic network of normal mammary epithelial cells is limited. Using a multi-omic approach, here we identify the diverse metabolic programmes operating in normal mammary populations. The proteomes of basal, luminal progenitor and mature luminal cell populations revealed enrichment of glycolysis in basal cells and of oxidative phosphorylation in luminal progenitors. Single-cell transcriptomes corroborated lineage-specific metabolic identities and additional intra-lineage heterogeneity. Mitochondrial form and function differed across lineages, with clonogenicity correlating with mitochondrial activity. Targeting oxidative phosphorylation and glycolysis with inhibitors exposed lineage-rooted metabolic vulnerabilities of mammary progenitors. Bioinformatics indicated breast cancer subtypes retain metabolic features of their putative cell of origin. Thus, lineage-rooted metabolic identities of normal mammary cells may underlie breast cancer metabolic heterogeneity and targeting these vulnerabilities could advance breast cancer therapy.

Press-fit fixation in anterior cruciate ligament reconstruction yields low graft failure and revision rates: a systematic review and meta-analysis.

PURPOSE: Press-fit fixation is a hardware-free technique in anterior cruciate ligament reconstruction (ACLR). The purpose of this review was to quantitatively assess the risk profile and outcomes of press-fit fixation and provide an update on its effectiveness compared to more standard fixation techniques of ACLR. METHODS: The electronic databases PUBMED, MEDLINE, and EMBASE were searched on March 26, 2020 for therapeutic randomized controlled trials (RCT) addressing press-fit fixation for primary ACLR. The Grading of Recommendations Assessment, Development and Evaluation tool was used to assess the quality for randomized studies. A meta-analysis with a random-effects model was used to pool applicable outcomes data. RESULTS: A total of six eligible RCTs were included in this review. There were 292 patients (72.9% male) with a mean age of 28.8 ± 3.8 years and a mean follow-up of 81.3 ± 88.3 months that underwent press-fit ACLR on the femoral, tibial or both tunnels. Femoral fixation techniques included press-fit fixation (96.6%) and cross-pin fixation (3.4%). Tibial fixation techniques included press-fit (37.0%), staples (28.1%), interference screws (21.2%) and abarticular post-screws (13.7%). Graft options included bone-patellar tend--bone autografts (73.6%) and semitendinosus and gracilis tendon autograft (26.4%). Significant improvements (p < 0.05) from baseline to follow-up were found for clinical outcomes. Significantly less postoperative bone tunnel enlargement (p < 0.05) was found with tibial press-fit fixation when compared to biodegradable screws. The overall complication rate was 13.3%. There were no significant differences in complication rates [odds ratio = 0.84 (95%CI 0.43-1.66); p = n.s.] (I2 = 0%) between patients undergoing femoral press-fit fixation and femoral metal interference screw fixation. CONCLUSION: The overall graft failure and revision rates with press-fit ACLR were low. There were no significant differences in complication rates between patients undergoing femoral press-fit and femoral metal interference screw fixation. Included studies found that patients undergoing press-fit fixation for ACLR had significant improvements in functional outcome scores postoperatively and had significantly reduced postoperative bone tunnel enlargement compared to patients undergoing bioabsorbable fixation. Thus, early evidence suggests that press-fit fixation appears to be a good option for patients undergoing ACLR. LEVEL OF EVIDENCE: I.

Mammary stem cells and progenitors: targeting the roots of breast cancer for prevention.

Breast cancer prevention is daunting, yet not an unsurmountable goal. Mammary stem and progenitors have been proposed as the cells-of-origin in breast cancer. Here, we present the concept of limiting these breast cancer precursors as a risk reduction approach in high-risk women. A wealth of information now exists for phenotypic and functional characterization of mammary stem and progenitor cells in mouse and human. Recent work has also revealed the hormonal regulation of stem/progenitor dynamics as well as intrinsic lineage distinctions between mammary epithelial populations. Leveraging these insights, molecular marker-guided chemoprevention is an achievable reality.

Identifying the murine mammary cell target of metformin exposure.

The heterogeneity of breast cancer makes current therapies challenging. Metformin, the anti-diabetic drug, has shown promising anti-cancer activities in epidemiological studies and breast cancer models. Yet, how metformin alters the normal adult breast tissue remains elusive. We demonstrate metformin intake at a clinically relevant dose impacts the hormone receptor positive (HR+) luminal cells in the normal murine mammary gland. Metformin decreases total cell number, progenitor capacity and specifically reduces DNA damage in normal HR+ luminal cells, decreases oxygen consumption rate and increases cell cycle length of luminal cells. HR+ luminal cells demonstrate the lowest levels of mitochondrial respiration and capacity to handle oxidative stress compared to the other fractions, suggesting their intrinsic susceptibility to long-term metformin exposure. Uncovering HR+ luminal cells in the normal mammary gland as the major cell target of metformin exposure could identify patients that would most benefit from repurposing this anti-diabetic drug for cancer prevention/therapy purposes.

A Novel 3-Dimensional Co-culture Method Reveals a Partial Mesenchymal to Epithelial Transition in Breast Cancer Cells Induced by Adipocytes.

Cancer metastases are accountable for almost 90% of all human cancer related deaths including from breast cancer (BC). Adipocytes can alter the tumor microenvironment, which can promote metastasis by inducing an epithelial-to-mesenchymal transition (EMT) in BC cells. However, the role of adipocytes during the mesenchymal-to-epithelial transition (MET), that can be important in metastasis, is not clear. To understand the effect of adipocytes on the BC progression, there is a requirement for a better in vitro 3-dimensional (3D) co-culture system that mimics the breast tissue and allows for more accurate analysis of EMT and MET. We developed a co-culture system to analyze the relationship of BC cells grown in a 3D culture with adipocytes. We found that adipocytes and adipocyte-derived conditioned media, but not pre-adipocytes, caused the mesenchymal MDA-MB-231 and Hs578t cells to form significantly more epithelial-like structures when compared to the typical stellate colonies formed in control 3D cultures. SUM159 cells and MCF7 cells had a less dramatic shift as they normally have more epithelial-like structure in 3D culture. Biomarker expression analysis revealed that adipocytes only induced a partial MET with proliferation unaffected. In addition, adipocytes had reduced lipid droplet size when co-cultured with BC cells. Thus, we found that physical interaction with adipocytes and ECM changes the mesenchymal phenotype of BC cells in a manner that could promote secondary tumor formation.