Breast Cancer Biopsies Research Articles

Polarization-dependent second-harmonic generation for collagen-based differentiation of breast cancer samples.

Nonlinear optical imaging techniques have been widely used to reveal biological structures for accurate diagnosis at the cellular as well as the tissue level. In the present study, polarization-dependent second-harmonic generation (PSHG) was used to determine collagen orientation in breast cancer biopsy tissues (grades 0, I, II and III). The obtained data were processed using fast Fourier transform (FFT) analysis, while second-harmonic generation (SHG) anisotropy and the "ratio parameter" values were also calculated. Such measurements were shown to be able to distinguish collagen structure modifications in different cancer grades tested. The analysis presented herein suggests that PSHG imaging could provide a quantitative evaluation of the tumor state and the distinction of malignant from benign breast tissues. The obtained results also allowed the development of a biophysical model, which can explain the aforementioned differentiations and is in agreement with the simulations relating the SHG anisotropy values with the mechanical tension applied to the collagen during cancer progression. The current approach could be a step forward for the development of new, nondestructive, label free optical diagnostic tools for cancer reducing the need of recalls and unnecessary biopsies, while potentially improving cancer detection rates.

Clinical study of exploring new axillary zone in sentinel lymph node biopsy of breast cancer

Objective: To explore the application of axillary reverse mapping (ARM) in breast cancer sentinel lymph node biopsy (SLNB), and to collect and record patient's data during operation. Through the specific experimental data, the anatomical location and morphology of the intercostal brachial nerve (ICBN) and the anatomic relationship of the axillary lymph nodes were analyzed to accurately locate the anatomical division of the axillary fossa of sentinel lymph node (SLN) and ARM lymph nodes. Methods: The technique of methylene blue staining for SLN combined with indocyanine green fluorescent staining for axillary reverse mapping was used to analyze the patients of Tianjin Central Hospital of Gynecology Obstetrics from June 2017 to June 2018. The clinical data of 35 patients with T1-2N0M0 breast cancer were analyzed. Results: Of the 35 patients, two cases were excluded from metastatic carcinoma of the SLN. Thirty-three cases were included in the data analysis. Three cases of ICBN were located 0-2.0 cm (9.09%) from the lower edge of the iliac vein, 27 cases were located at 2.0-4.0 cm (81.82%), and 3 cases of ICBN were located greater than 4.0 cm (9.09%). In the region of 0-2.0 cm from the lower edge of the iliac vein, 1-2 ARM lymph nodes were found in 5 cases; in the 2.0-4.0 cm area, SLN was found in 33 cases, 4 of which found 1 coincident lymph node; at>4.0 cm ARM lymph nodes and overlapping lymph nodes were not found in the cm region, and 1-2 SLNs were found in 3 cases. Conclusions: Individual ICBNs vary greatly, with different shapes and distances from the lower edge of the axillary vein. The axillary region below the iliac vein can be divided into three regions: ARM region (0-2.0 cm); SLN region (2.0-4.0 cm); and sentinel gate region (>4.0 cm). The position 2.0 cm from the lower edge of the iliac vein can be used as the boundary between the ARM lymph node and the SLN. 4. There is a certain proportion of coincidence rate between SLN and ARM lymph nodes.

IL-31 induces antitumor immunity in breast carcinoma

BackgroundImmunomodulatory agents that induce antitumor immunity have great potential for treatment of cancer. We have previously shown that interleukin (IL)-31, a proinflammatory cytokine from the IL-6 family, acts as an...

Wide local excision and sentinel lymph node biopsy in early breast cancer in Duhok city

Wide local excision and sentinel lymph node biopsy in early breast cancer in Duhok city

Clinical Correlations of Programmed Cell Death Ligand 1 Status in Liquid and Standard Biopsies in Breast Cancer.

Data regarding the prognostic value of programmed cell death ligand 1 (PD-L1) expression on circulating tumor cells (CTCs) are lacking. However, CTCs could represent an alternative approach to serial biopsies, allowing real-time monitoring of cancer phenotype. We evaluated, in a dedicated prospective clinical trial, the clinicopathological correlations and prognostic value of PD-L1(+)-CTCs in 72 patients with metastatic breast cancer (MBC). Eighteen of 56 patients with available archival tissue presented at least one positive (≥1%) PD-L1 tumor sample. Baseline CTCs and PD-L1(+)-CTCs were detected in 57 (79.2%) and 26 (36.1%) patients. No significant correlation was found between PD-L1 tumors and CTC expression. In univariate analysis, triple negative (TN) phenotype, number of metastatic treatments, >2 metastatic sites, ≥5 CTCs and PD-L1(+)-CTCs were significantly associated with progression-free survival, while tissue PD-L1 expression was not. In multivariate analysis, TN phenotype, number of metastatic treatments and of metastatic sites were the only 3 variables independently associated with progression-free survival. Progesterone receptor negativity, TN phenotype, >2 metastatic sites and ≥5 CTCs were significantly associated with overall survival in univariate analysis. In multivariable analysis, TN phenotype and >2 metastatic sites were the only 2 independent variables. Unlike PD-L1(+)-tumor, PD-L1(+)-CTCs correlate to survival in MBC. Reappraisal of the role of PD-L1 expression by tumor tissue and by CTCs under anti-PD-1/PD-L1 treatment is necessary to evaluate its predictive value and potential role as a stratifying factor in strategies and trials for MBC patients with MBC. NCT02866149.

Preoperative tumor biopsy results in more detected sentinel nodes than intraoperative biopsy in breast cancer patients

BackgroundSentinel lymph node biopsy (SLNB) plays a vital role in breast cancer surgery, and the identified number of sentinel nodes determines its accuracy for representing the status of the axillae. There are two types of tumor biopsies in breast cancer: preoperative and intraoperative biopsies. We compared the effects of the two different biopsies on the results of SLNB.MethodsPatients with clinical stages T1–3, N0 (cT1-3 N0) tumors were enrolled in this study. A total of 53% of patients received preoperative tumor biopsy, and 47% received intraoperative excisional biopsy. To identify the sentinel lymph nodes, patients received dual tracer injection. The number of SLNs detected and the false-negative rate were compared between groups.ResultsA total of 204 patients were enrolled, 108 received preoperative tumor biopsy, and 96 received intraoperative excisional biopsy. Among all the patients, 160 received axillary lymph node dissection (ALND) following SLNB. Preoperative tumor biopsy detected more SLNs than intraoperative biopsy (mean rank 113.87 vs. 90.9, p = 0.004). The false-negative rates in the preoperative and intraoperative tumor biopsy groups were 3% and 18%, respectively.ConclusionsPatients in the preoperative tumor biopsy group had more SLNs identified than intraoperative biopsy patients. The false-negative rate was also lower in the preoperative biopsy group.

One-step nucleic acid amplification CK19 copy number for sentinel node biopsy in breast cancer: Identification of new cutoffs to predict nonsentinel axillary node involvement.

We evaluated novel sentinel node values in breast cancer for one-step nucleic acid amplification (OSNA) to predict further nodal involvement using various methods in clinically node-negative disease with a positive OSNA result and subsequent axillary node dissection. 239 patients (118 macrometastatic) were assessed revealing cutoffs of total tumor load (TTL) 44500copies/µL (AUROC 0.793); average copy number (ACN) 9450 (AUROC 0.790); and highest copy number (HCN) 46,000. For macrometastatic patients only: TTL 221 400copies/µL (AUROC 0.685); ACN 64,000 (AUROC 0.671); HCN 59 500 (AUROC 0.529). Our data favor TTL and represent one of the largest OSNA macrometastatic predictive series.

Superparamagnetic iron oxide: a novel tracer for sentinel lymph node detection in vulvar cancer

ObjectivesSuperparamagnetic techniques for sentinel lymph node (SLNs) biopsy in breast cancer is well recognized but remains novel in the literature in relation to early stage vulvar cancer. The aim of...

ASO Author Reflections: The De-Escalation of Sentinel Node Biopsy for Breast Cancer

ASO Author Reflections: The De-Escalation of Sentinel Node Biopsy for Breast Cancer

ASO Author Reflections: Core Needle Biopsy and Hormonal Receptor Retesting in Breast Cancer: Controversy and Management.

ASO Author Reflections: Core Needle Biopsy and Hormonal Receptor Retesting in Breast Cancer: Controversy and Management.

A Pilot Study for the Feasibility of Exome-Sequencing in Circulating Tumor Cells Versus Single Metastatic Biopsies in Breast Cancer.

The comparison of the landscape of somatic alterations in circulating tumor cells (CTCs) versus metastases is challenging. Here, we comprehensively characterized the somatic landscape in bulk (amplified and non-amplified), spike-in breast cancer cells, CTCs, and metastases from breast cancer patients using whole-exome sequencing (WES). We determined the level of genomic concordance for somatic nucleotide variants (SNVs), copy number alterations (CNAs), and structural variants (SVs). The variant allele fractions (VAFs) of somatic variants were remarkably similar between amplified and non-amplified cell line samples as technical replicates. In clinical samples, a significant fraction of somatic variants had low VAFs in CTCs compared to metastases. The most frequently recurrent gene mutations in clinical samples were associated with an elevated C > T mutational signature. We found complex rearrangement patterns including intra- and inter-chromosomal rearrangements, singleton, and recurrent gene fusions, and tandem duplications. We observed high molecular discordance for somatic alterations between paired samples consistent with marked heterogeneity of the somatic landscape. The most prevalent copy number calls were focal deletion events in CTCs and metastases. Our results demonstrate the feasibility of an integrated workflow for the identification of a complete repertoire of somatic alterations and highlight the intrapatient genomic differences that occur between CTCs and metastases.

Comparison of clinical outcomes between sentinel lymph node biopsy and axillary lymph node dissection in a single-center Z0011-eligible breast cancer cohort.

The ACOSOG Z0011 trial has proven the oncological safety of sentinel lymph node biopsy (SLBx) for node negative breast cancer. Accordingly, treatment paradigm including axilla surgery was changed. We retrospectively reviewed breast cancer patients to evaluate the clinical effect of paradigm shift in breast cancer surgery after applying the Z0011 criteria. All women who underwent breast-conserving surgery at the National Cancer Center between January 1, 2000, and December 31, 2015, were enrolled and classified according to the Z0011 criteria. The primary endpoint of the study was the disease-free survival rates, and the secondary was the adverse events, especially arm lymphedema. Total 361 patients were enrolled the study (271 axillary lymph node dissection [ALND] group, 90 SLBx group). After the Z0011 guideline was adopted in our institute, the use of ALND decreased, and lymph node sampling (removing only a few axillary lymph nodes) replaced ALND. The total mean number of retrieved nodes were more in ALND group (13.02) than SLBx group (3.43). However, there was no difference in the mean number of positive nodes between two groups (2.34 in ALND group vs. 1.12 in SLBx group, P=0.001). During follow-up, 25 patients experienced disease recurrence: 22 from the ALND group and three from the SLBx group. All of died seven patients were from the ALND group. The ALND group had more complications than the SLBx group (P=0.02). Arm edema occurred more frequently in the ALND group (29.5%) than in the SLBx group (5.6%), although without statistical significance (P=0.07). In our study, we concluded that SLBx can be used safely in Z0011-eligible cohort without increased risk of locoregional recurrence. Moreover, we found that omission of ALND is favored to reduce some serious complications such as arm lymphedema.

A randomized study comparing different doses of superparamagnetic iron oxide tracer for sentinel lymph node biopsy in breast cancer: The SUNRISE study

IntroductionThe non-radioactive method that uses the magnetic tracer (SPIO/Sienna) has shown to be a feasible technique for the SLN detection in breast cancer patients. The aim of this study is to assess the efficacy of different doses of a new magnetic tracer Sienna XP (Magtrace) compared to Tc-99 m and to evaluate its non-inferiority. MethodsPatients diagnosed with early-stage breast cancer cT1-3 N0, from October 2016 to August 2018 were eligible and consecutively randomized to three different doses of new SPIO used: group 1 (1 mL), group 2 (1.5 mL) and group 3 (2 mL). ResultsA total of 135 patients were included in the study, 45 in each group. Detection of SLNs with the three doses of Sienna XP (1 mL, 1.5 mL and 2 mL) showed non-inferior rates compared to the conventional technique with radiotracer (p = 0.654). Concordance by patients with SLN positive was 100% for all groups.83 (70.3%) patients reported skin staining at one month postoperatively, significantly lower in group 1 (p = 0.042). At 6 months follow up, group 1 remains with significantly lower skin discoloration (p = 0,01). In multivariate analysis, dose of 2 mL showed statistically significant for the skin staining. The majority of patients (70%) felt that skin discoloration does not represent a problem. ConclusionThe use of the Sienna XP magnetic tracer at 1 mL is not inferior to higher doses of magnetic tracer neither is inferior to radiotracer. 1 mL of magnetic tracer resulted in significantly less skin discoloration compared to higher doses.

Circulating Tumor Cell and Regulator T-Lymphocyte in Core Biopsy for Breast Cancer

Introduction: Breast cancer is the most common malignancy found in Indonesia. Core biopsy is one of the modalities used in breast cancer diagnosis with sensitivity of 91-99% and specificity of 96-100%. The procedure causes damage to tumor tissue thus causing tumor cells to enter circulation (CTC) and therefore acute inflammation and infiltration of inflammatory cells. In the final phase of infiltration, the number of Tregs cells will increase, as well as secretion of TGFβ and IL-10, creation of immunosuppresion microenvironment, COX2 stimulation by TGFβ then conversion of CD4+ T cells into FoxP3+ (Tregs), therefore the number of Tregs cells will increase. SOX-4 is activated by TGFβ, and then EMT proccess occurs, tumor cells enter circulation and CTC number will increase. Considering the side effect of core biopsy which is entrance of tumor cells to circulation causing the procedure to be in debate/contradicting opinions. Based on this premise, this study aims to investigate whether there is a change and relationship in Tregs count and CTC count before and after core biopsy procedure.

The Diagnostic Significance of Ultrasound and Ultrasound-guided Coarse Needle Biopsy in Breast Cancer in Pregnancy

Objective: To explore the significance of ultrasound and ultrasound-guided coarse needle biopsy in the clinical diagnosis of breast cancer patients in pregnancy. Methods: A total of 10 patients with breast cancer in pregnancy were selected to study from October 2018 to October 2019 in our hospital, who were given preoperative ultrasound results and ultrasound-guided coarse-needle biopsy histopathology, and the results were analyzed. Results: All the 10 patients showed thickening of the mammary gland body, appeared to pick up disorderly situation, most of the dilated catheter, including 8 patients with solid hypoechoic mass, irregular shape, and "crab foot" edge burr. The corresponding aspect ratio value was greater than 1. The other 2 patients presented with cystic mixed masses with poor boundary definition, irregular shape, and mostly accompanied by posterior echo enhancement. Conclusion: Simple ultrasound diagnosis for patients with breast cancer during pregnancy has some errors, and should be used in combination with ultrasound-guided coarse needle biopsy operation. This method has a high clinical diagnosis rate and significant clinical application value, so it is worth promoting.

Methylene Blue Absorption in Sentinel Lymph Node Biopsy for Early Breast Cancer after Neoadjuvant Chemotherapy.

Introduction:Chemotherapy is claimed to cause lymphatic drainage damage because of the tumor cell’s apoptosis process. This event might cause decreased marker (radioactive solution and/or blue dye) absorption on sentinel lymph nodes (SLN). In this study, the researchers used methylene blue only and wished to evaluate the methylene blue absorption of the SLNB procedure on early-stage breast-cancer patients after neoadjuvant chemotherapy (NAC). Materials and methods:The method used was the historical cohort study conducted from 2016-2019 in Indonesia. Samples were collected from 117 patients of stage I and II breast cancer with clinically negative axillary lymph nodes, who were then grouped into post-NAC and no-NAC (control group), in which SLNB procedures were conducted on the two groups by using single-method methylene blue. The results of methylene blue absorption were then analyzed by the Chi-square hypothesis test. Results:From the total of 564 early-stage patients who were referred to surgical oncologists, 117 patients were found to meet criteria of inclusion, consisting of the control group (52 patients) and the post-NAC group (65 patents). Of 65 patients who had undergone NAC treatment and SLNB procedure, it was found that 40 patients (61.5%) showed positive blue SLN. Of 52 pre-NAC breast-cancer patients, it was found that 47 patients (90.4%) showed methylene blue absorption on SLN with the p-value of 0.000 (P<0.05, significant). The relative risk value amounted to 0.522. Post-NAC patients had a tendency of decreased absorption of methylene blue. Conclusion:Neoadjuvant chemotherapy can cause the decrease of methylene blue absorption on SLNB procedure.

Abstract B124: Breast cancer treatment delays at an urban safety net hospital among women experiencing homelessness

Abstract Abstract Disparities in outcomes for vulnerable women is a persistent, ongoing problem. Timely treatment improves breast cancer outcomes and efforts to improve delays among underserved patients is needed. Specifically, homelessness and breast cancer treatment outcomes are understudied. This is a novel and descriptive study exploring types of homelessness and treatment delays at an urban safety net hospital providing care to a vulnerable patient population. Experimental Procedures This study is a retrospective chart review of homeless female patients diagnosed with breast cancer between January 1, 2000 and December 31, 2014. Data for this study were acquired from the hospital cancer registry and electronic medical record. Homelessness was categorized as transitionally, episodically or chronically homeless. Other variables collected included demographic characteristics and time to treatment. A detailed chart review was conducted to identify delays to breast cancer treatment and the potential reasons for delay between diagnosis and first treatment. All demographic characteristics, time to treatment and factors related to delays to treatment were analyzed descriptively, reporting frequencies and proportions. Delay to treatment was calculated as date of pathologically confirmed biopsy of breast cancer to date of first treatment (surgery or chemotherapy). Summary of Data The total number of individuals analyzed was 24. All except two subjects were delayed to treatment (&amp;gt; 30 days from diagnosis to treatment). Most women in this cohort were categorized as chronically homeless (46%) with the rest categorized as transitionally (29%) or episodically (12%) homeless. The majority of subjects (70%) were Black, non-Hispanic. Most women identified as single (58%) or divorced (21%) at the time of breast cancer diagnosis. All except one subject were publicly insured (71% Medicaid; 12% Medicare) or uninsured (8%). Regardless of type of homelessness, most subjects were either 30-60 or 60-90 days delayed. Those who were chronically homeless experienced significantly more delays to first treatment (56% of those who were delayed 30-60 days and 57% of those who were delayed 60-90 days; p-value 0.006) than those who were episodically or transitionally homeless. Conclusions Significant delays and barriers to breast cancer treatment exist among women experiencing homelessness. Further studies and focused efforts to improve timely breast cancer care for homeless women is warranted. Citation Format: Kate Festa, Ariel E Hirsch, Michael R Cassidy, Lauren Oshry, Kathryn Quinn, Margaret M Sullivan, Naomi Y Ko. Breast cancer treatment delays at an urban safety net hospital among women experiencing homelessness [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr B124.

Abstract D063: Intratumoral macrophage analysis and characterization of HER2+ pretreatment breast tumor biopsy using multiplex immunofluorescence

Abstract Background: Latinas in the US experience overall worse breast cancer (BC) survival than non-Latinas. Higher rates of aggressive BC subtypes including estrogen receptor negative(ER-) and HER2+ in Latinas are partly suspected. High intra-tumoral immune cells including myeloid suppressive M2 type macrophages (MAC) that suppress tumor immune responses are positively associated with poor outcomes and low response to therapy. Unclear is if tumor associated immune cells differ by race/ethnicity. Objective: To develop and evaluate the performance of a multiplex immunofluorescence (IF) methods to classify tumor-associated macrophages (TAM) as M1 and M2 in HER2+ breast tumors. Methods: Two panels of three antibodies were studied in HER2+ breast cancer biopsy specimens (n=19 patients). Two myeloid panels targeting CD80, CD68, CD163, CD206 with a nuclear (DAPI) and tumor cell (cytokeratin 8/18) was investigated. Banked breast tumor tissues were deparaffinized, and stained using indirect multiplex IF. Each tissue underwent multiple staining for CD206, CD68, and CD163 with stripping. A tyramide signal amplification IF (Perkin Elmer) was used in fluorescent labeling. Whole slide analysis image analysis was performed using HALO image analysis software. Results: All biopsy tissues yielded high quality IF profiles. Analyses per patient tumor (i.e., 5 regions of high tumor cell density containing a minimum of 5000 nucleated cells) revealed a fairly homogeneous MAC profile. In contrast, significant between patient heterogeneity in intratumoral M2 MAC density and differences by ER status was observed. In this sample, Her2+ BCs contained a high intratumoral TAM content – i.e., mean 27% (range &amp;lt;1% to 70%) of cells in tumor dense regions were positive for the pan MAC marker CD68+. Further, intratumoral CD68+ cells (all MAC) were slightly higher in abundance in ER-/HER2+ tumors at 31% compared to 20% in ER+/HER2+ tumors. Further, the relative abundance of CD68+/CD206+ cells (M2 type) in tumor rich regions showed a high degree of variability between cases with a mean 7.1% of total cells (range 0 to 48%). Notable, the relative abundance of double CD68/CD206 cells was higher in the ER+ tumors at 8.8% (range 0 to 48%) compared to 3.3% in ER- that showed a narrow range of 0 to 8%. A similar pattern was observed for the CD68/CD163 double positive M2 type that included large heterogeneity between patients and higher abundance in ER+/HER2+ tumors. In contrast, while exhibiting high heterogeneity between patients, the triple positive CD68, CD163, CD206 cells population appeared to be proportionally higher in ER-/HER2 tumors. Ongoing work includes characterization of the M1 type MAC and their relative abundance in HER2+ tumors by ER status. Conclusion: These results demonstrate the feasibility of multiplex IF to characterize TAM subsets in patient derived BC tissues. Future efforts will include relating TAM to HER2 targeted therapy response rates and examination of differences in TAM profiles in patient populations by race/ethnicity. Citation Format: Emmaly D Gutierrez, Christina Preece, Alison Stopeck, Patricia Thompson. Intratumoral macrophage analysis and characterization of HER2+ pretreatment breast tumor biopsy using multiplex immunofluorescence [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr D063.

Detection of actionable gene mutations in breast cancer by amplicon-based next-generation sequencing liquid biopsy.

1035 Background: The PI3K-inhibitor, alpelisib, was approved for the treatment of hormone receptor (HR)-positive, HER2-negative breast cancers with PIK3CA mutations based on findings from the SOLAR-1 trial. PIK3CA-positivity in tumor tissue was 29% (341/1172), but was only 15% (177/1172) in plasma cell-free DNA (cfDNA). The lower mutation detection rate observed in cfDNA may limit the clinical application of liquid biopsy in breast cancer. Amplicon-based next-generation sequencing (NGS) approach may confer improved sensitivity, allowing more effective profiling. We conducted a study to evaluate the use of this technology. Methods: Plasma cfDNA from 113 breast cancer patients (82.3% metastatic) underwent real-world testing in a CAP and ISO15189 accredited central laboratory. We analysed genetic alterations in cfDNA using an amplicon-based NGS technology. The presence of PIK3CA and other mutations relevant to breast cancer were correlated to molecular subtypes and treatment histories. Results: At least one mutation was detected in 70.8% of cases. Mutations were more frequent in metastatic cases (77.4%) compared to non-metastatic cases (27.3%). Across all patients, mutations in PIK3CA (33.6%), TP53 (32.7%), ESR1 (22.1%), GATA3 (7.1%) and ERBB2 (7.1%) were most frequently detected, in accordance with tumor tissue genotyping studies. PIK3CA mutations were more common in HR+ HER2- patients (44.4% vs 28.6% of other patients). Among PIK3CA-mutant cases, multiple PIK3CA mutations were present in 18.4% of cases, and hotspot mutations H1047R (34.2%), E542K (26.3%) and E545K (15.8%) were most frequent. An association was seen between PIK3CA mutation and prior treatment with CDK4/6 inhibitors (palbociclib, ribociclib) or mTOR inhibitor (everolimus), with 58% of PIK3CA-mutant cases having received these treatment previously compared to only 20% of PIK3CA-wild type (wt) cases. In addition, 75% of previously treated ESR1-mutant cases had specifically received hormonal treatment, compared to 60 % of ESR1-wt cases that received any treatment. Conclusions: We report similar PIK3CA mutation frequencies (~30%) with amplicon-based NGS on cfDNA compared to tumor tissue testing in breast cancer. Importantly, other driver mutations were also observed at similar frequencies as external tissue studies, implying high sensitivity as the primary reason for performance. This supports the clinical utility of an amplicon-based NGS-based approach to liquid biopsy for the sensitive detection of actionable mutations in breast cancer.

Presence of human papillomavirus DNA in breast cancer patients: SERUM and tissue analysis.

e12582 Background: It has been demonstrated an increased risk of breast cancer (BC) incidence in patients with previous cervical dysplasia, suggesting a role of human papillomavirus (HPV) of cervical lesions in the development of BC. Although, the origin of HPV spreading to the mammary gland and its mechanism of dissemination is not clear. Methods: Seven serum samples from healthy donors and 58 from early BC patients collected pre-surgery were analyzed for the presence of HPV DNA. For 49/58 patients HPV DNA was analyzed also on the primary tumor tissue. 17 patients had luminal A BC (4 relapsed, 13 non relapsed), 16 had luminal B BC (5 relapsed, 11 non relapsed), 13 had triple-negative BC (6 relapsed, 7 non relapsed), 12 had HER2-positive BC (4 relapsed, 8 non relapsed). Circulating DNA was extracted from 500 μl of serum by Qiamp DNA Mini kit (Qiagen, Milan, Italy) and tumor DNA was extracted from at least four 10-micron sections by QIAamp DNA FFPE Tissue Kit (Qiagen, Milan, Italy). Circulating HPV DNA was amplified by a multiplex PCR with HPV E6 or E7 gene-specific primers and the sequence was assessed by a high-throughput MALDI-TOF mass spectrometry-based method. Results: HPV DNA was detected in only 5 serum of BC patients and in none of the healthy controls. 4/5 BC cases had high-risk HPV DNA (type 39,45,52,59) and 1 had type 73 low-risk HPV DNA. 4/5 HPV DNA-positive patients had previously low-grade cervical intraepithelial neoplasia (CIN I) detected by Pap smear. These 5 patients with circulating HPV DNA did not show HPV positivity in the BC tissue. 2 out of 49 cases were positive for universal HPV DNA sequence in tissue and only 1 case showed HPV type 51. No relation was found between HPV infection and tumor subtype or prognosis, neither for HPV DNA positivity between serum and tissue. Conclusions: Our data support the feasibility of HPV DNA detection by liquid biopsy in BC. The presence of circulating HPV could be due to a viral spread from other organs. More data are needed to establish the role of circulating HPV DNA and its potential association with HPV infection of the breast and/or of the cervix. [Table: see text]