bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P1-06-05: Breast cancer organoid cultures preserve intra-tumor heterogeneity and reveal intrinsically resistant phenotypes to standard chemotherapies
Authors: Sowder ME , Ludwik KA , Pasic L , Brenin DR , Stricker TP , Macara IG , Lannigan DA 
Published: March 2017

Abstract
Breast cancer intra-tumor heterogeneity contributes to chemotherapy resistance and decreased patient survival, yet no reliable in vitro models exist to study this phenomenon. To address this need we developed an in vitro 3D organoid culture system using primary human breast cancer tissue. A major difficulty in the development of such models is to identify robust in vitro conditions that preserve the breast cancer phenotypes observed in situ. To address this challenge we used quantitative immunofluorescence imaging to compare the cellular phenotypes in the starting tumor tissue with those observed in the tumor organoids cultured in 3D. We utilized a clustering algorithm and utility function to quantitatively assess whether tumor organoids generated in vitro faithfully recapitulated intra- and inter-tumor heterogeneity of the tumor tissue in situ. This approach generated a normalized score that reflects tissue-organoid similarity. To test the sensitivity of our method to overall changes in tissue phenotype we focused on three distinct breast cancer subtypes distinguished by expression of estrogen receptor (ER), progesterone receptor (PR) and amplification of ERBB2/HER2 (HER2). Using our approach, we successfully recapitulated the tumor phenotypes present in ER+, ER+/HER2+ and triple negative breast cancer. We discovered that EGF preserves the TNBC phenotype, whereas AREG is required for recapitulating the phenotype of ER+ and ER+/HER2+ breast cancers. Additionally, our data demonstrate that HER1 ligands drive inter- and intra-tumor heterogeneity. To investigate how intra-tumor heterogeneity contributes to therapy responses we treated organoids with standard agents used clinically to treat each of the distinct subtypes. For all tumor subtypes we observed differential vulnerabilities between patients to drug treatments. Importantly, our analysis identified divergent cellular phenotypes that have various sensitivities to chemotherapies. Taken together, our methodology provides an unprecedented view of intra-tumor heterogeneity and allows for the investigation of chemo-resistance mechanisms. Further, this approach will provide a powerful tool, which will enhance the identification of novel therapies and facilitate personalized medicine.
Citation Format: Sowder ME, Ludwik KA, Pasic L, Brenin DR, Stricker TP, Macara IG, Lannigan DA. Breast cancer organoid cultures preserve intra-tumor heterogeneity and reveal intrinsically resistant phenotypes to standard chemotherapies [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-06-05.
Abstract P1-06-06: Evidence for tumor heterogeneity and clonal evolution during invasive progression of breast cancer
Authors: Ding Y , Marks JR , King LM , Hall AH , Mardis ER , Rodrigo AG , Maley CC , Hwang E-S 
Published: March 2017

Abstract
Purpose: Intratumoral heterogeneity is well recognized to be an important driver of treatment resistance and metastasis. We undertook this N of one study to measure the degree of heterogeneity in a single large preinvasive lesion with an invasive component to determine the relationship between tumor heterogeneity, spatial distribution, clonal evolution, and invasive progression.
Methods: We identified a patient with extensive DCIS measuring 7.5 cm, associated with 1.5 cm of an invasive component. We segregated the tumor sample into 32 unique blocks with precise geospatial localization; invasive cancer was identified in 3 of 32 blocks. NGS libraries were made from FFPE derived DNA (20-40ng) for full exome sequencing and hybridization to a 4.8 million element SNP array. All data were analyzed and a phylogentic tree was constructed.
Results: The sequence data was analyzed with Platypus[1] and 3922 somatic mutation sites were found in total. These sites were concatenated into one sequence for each sample. Then a Neighbor-Joining tree was built with a Jukes-Cantor model and 1000 bootstrap replicates using FastME 2.0[2], to assess the reliability of the tree (Figure 1). Phylogenetic analysis revealed that invasive cancer evolved twice, independently. Dense sampling allowed reconstruction of the temporal order of mutations that accumulated in the cell lineage of the invasive cancers. Furthermore, the phylogeny revealed that distant regions may be closely genetically related, the oldest parts of the tumor were in the interior of the tumor, and that the invasive tumors evolved near the oldest parts of the tumor, rather than at the expanding front.
Conclusions: Extensive sampling and sequencing of a single tumor yields important insights about tumor heterogeneity and tumor progression for DCIS to invasive cancer. Foci of invasion were geospatially associated with preinvasive regions of progressively higher mutational load.
1. Rimmer A, Phan H, Mathieson I, Iqbal Z, Twigg SR, Wilkie AO, McVean G, Lunter G, Consortium W: Integrating mapping-, assembly-and haplotype-based approaches for calling variants in clinical sequencing applications. Nature genetics 2014, 46(8):912-918.
2. Lefort V, Desper R, Gascuel O: FastME 2.0: a comprehensive, accurate, and fast distance-based phylogeny inference program. Molecular biology and evolution 2015:msv150.
Citation Format: Ding Y, Marks JR, King LM, Hall AH, Mardis ER, Rodrigo AG, Maley CC, Hwang E-S. Evidence for tumor heterogeneity and clonal evolution during invasive progression of breast cancer [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-06-06.
Abstract P1-08-07: Assessing the impact of loss of NF1 protein on endocrine therapy resistance
Authors: Cakar B , Chan D , Yan P , Zheng Z , Singh P , Lei JT , Haricharan S , Ellis M , Chang E 
Published: March 2017

Abstract
Background: The vast majority of breast cancers belong to the luminal subtype, which expresses the estrogen receptor-α (ER). Although great strides have been made in targeting the ER pathway for treating the ER+ tumors, relapse and death is common and ongoing. In order to identify the cause for treatment resistance, we have conducted a retrospective analyses on the tumor ge­nomes of &gt;600 patients treated by tamoxifen monotherapy in the adjuvant setting with a median follow-up of 10.4 years. Our data have revealed that NF1 (Neuro­fibromatosis type 1) gene loss of function mutations were greatly associated with poor prognosis. NF1 is a tumor suppressor acting mostly as a GAP (GTP ase activating protein) to switch off acti­vated Ras. We aim to define the impact of loss of NF1 protein on patient outcome in ER+ breast cancer patients by establishing an immunohistochemistry (IHC) protocol to detect NF1.
Method and results: We have first surveyed commercially available antibodies by Western blot and found one that could efficiently detect endogenous NF1. We then use this to validate inducible shRNA clones against NF1, as well as a breast cancer cell line that is NF1-null. This antibody has high background. We have thus partially purified a commercially available NF1 antibody by preclearing using NF1-null cell lysate. We then performed immunostaining using NF1-silenced and null cells as control and found that NF1 is mostly cytoplasmic and nuclear. To get antibody of high quality, we have decided to make our own antibody by expressing a C-terminal fragment of NF1 as a GST-tagged protein (GST-NF1c). Production of polyclonal and monoclonal antibody is in progress.
Conclusion: Our clinical profiling data suggest that loss of NF1 protein, a very common event in a wide range of other cancers, promotes endocrine therapy resistance. An efficient IHC protocol will enable us to firmly validate whether loss of the NF1 protein indeed correlates with poor patient outcome. This method will ultimately enable us to identify high risk NF1 deficient patients and to properly treat them.
Citation Format: Cakar B, Chan D, Yan P, Zheng Z, Singh P, Lei JT, Haricharan S, Ellis M, Chang E. Assessing the impact of loss of NF1 protein on endocrine therapy resistance [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-08-07.
Abstract P1-10-09: Delineation of internal mammary nodal target volumes in breast cancer radiotherapy
Authors: Jethwa KR , Kahila MM , Hunt KN , Brown LC , Corbin KS , Park SS , Yan ES , Boughey JC , Mutter RW 
Published: March 2017

Abstract
Purpose/Objectives: The optimal clinical target volume (CTV) for internal mammary node irradiation (IMNI) is uncertain in an era of increasingly conformal volume-based treatment planning for breast cancer. We mapped the location of gross internal mammary lymph node (IMN) metastases in order to identify areas at highest risk for harboring occult disease.
Methods and Materials: Patients with axial imaging of IMN disease, including fluorodeoxyglucose F-18 (FDG18) positron emission tomography (PET-CT) and magnetic resonance imaging (MRI), were identified from a breast cancer registry. The IMN location was transferred by a radiation oncologist and breast radiologist onto the corresponding anatomic position on representative axial CT images of a patient in the treatment position. Distribution of lymph nodes, and their location was compared with consensus group guidelines of IMN target delineation.
Results: Sixty-seven patients with 130 IMN metastases were mapped. The location was in the first three intercostal spaces in 102 of 130 (78%) nodal metastases. Eighteen of 130 (14%) IMN were located caudal to the third intercostal space, while 10 of 130 (8%) IMN were located cranial to the first intercostal space, including 3 patients with isolated IMN metastases at that location in the absence of distant disease. Of the 102 nodal metastases within the first three intercostal spaces, 54 (53%) were located within the RTOG consensus volume. Relative to the internal mammary (IM) vessels, 19 (19%) nodal metastases were located medially with a mean distance of 2.2 mm (SD 2.9 mm), while 29 (28%) were located laterally with a mean distance of 3.6 mm (SD 2.5 mm). Ninety and ninety-five percent of lymph nodes within the first three intercostal spaces would have been encompassed within a 4 mm and 6 mm medial and lateral expansion on the IM vessels, respectively.
Conclusions: For women with indications for elective IMNI, a 4 mm medial and lateral expansion on the IM vessels within the first 3 intercostal spaces may be appropriate. In women with known IMN involvement, cranial extension to the confluence of the IM vein with the brachiocephalic vein +/- caudal extension to the 4th or 5th interspace with a 6 mm medial and lateral expansion may be considered, provided that normal tissue constraints are met.
Citation Format: Jethwa KR, Kahila MM, Hunt KN, Brown LC, Corbin KS, Park SS, Yan ES, Boughey JC, Mutter RW. Delineation of internal mammary nodal target volumes in breast cancer radiotherapy [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-10-09.
Abstract P1-10-17: Radiotherapy associated with concurrent bevacizumab in patients with non-metastatic breast cancer
Authors: Dautruche A , Belin L , Cottu P , Bontemps P , Lemanski C , De La Lande B , Baumann P , Missohou F , Levy C , Peignaux K , Reynaud-Bougnoux A , Denis F , Gobillion A , Ady Vago N , Fourquet A , Kirova Y 
Published: March 2017

Abstract
Purpose/Objectives
The purpose of this study was to determine early and late toxicities among patients with non-metastatic breast cancer (BC) receiving concurrent bevacizumab (BV) and radiation therapy (RT).
Materials/Methods
Multicentre, prospective study, of the toxicity of adjuvant concomitant association of BV and RT in patients with non-metastatic BC enrolled in Phase 3 BEATRICE, BEVERLY and BETH trial. Early and late toxicities were assessed by the Common Terminology Criteria for Adverse Events v. 3.0 during RT, 12 months and 36 months after its completion.
Results
Sixty-four patients were included from october 2007 to august 2010. They all received adjuvant RT and BV concomitant treatment, plus neo-adjuvant BV for 24 patients. RT was adjuvant and normo-fractionated. Twelve months toxicity was available for 60 patients and 36 months toxicity was available for 43 patients. Median follow-up was 46 months (18-77). Median age was 51 years old (23-68). Among 63 evaluated patients during RT, acute radiation dermatitis was observed in 48 (76%) patients : Grade 1 for 27 (43%), grade 2 for 17 (27%), grade 3 for 4 patients (6%). Grade 2 acute oesophagitis was observed in 1 patient. At 3 years, few toxicities were observed : 6 patients (14%) had grade 1 pain, 4 (9%) had grade 1 fibrosis, one (2%) had grade 1 telangiectasis, one (2%) had grade 1 paresis, 3 (7%) had grade 1 lymphoedema and one grade 3 lymphoedema. No grade 4 toxicity was observed. At 12 months, only one evaluated patient had a LVEF &lt;50% and none at 36 months.
Conclusions
Concurrent bevacizumab with locoregional RT is associated with acceptable early and late 3-years toxicities in patients with BC. Determination of late toxicity at 60 months is currently underway.
Citation Format: Dautruche A, Belin L, Cottu P, Bontemps P, Lemanski C, De La Lande B, Baumann P, Missohou F, Levy C, Peignaux K, Reynaud-Bougnoux A, Denis F, Gobillion A, Ady Vago N, Fourquet A, Kirova Y. Radiotherapy associated with concurrent bevacizumab in patients with non-metastatic breast cancer [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-10-17.
Abstract P1-11-10: Clinical applications of near infra-red imaging system for localization of non-palpable breast lesions in breast conserving surgery
Authors: Jung S-Y , Lee S , Kim SK 
Published: March 2017

Abstract
Backgrounds: Localization of non-palpable breast lesions is important for obtaining tumor-free resection margin and achieving better cosmetic outcome. Near infra-red (NIR) imaging system has been introduced for localization in breast surgery. This study aimed to evaluate the feasibility of localization using NIR imaging system in breast conserving surgery (BCS) (ClinicalTrials.gov identifier: NCT 02172989/ NCT02473159).
Materials and Methods: Between June 2014 and October 2015, 20 women with benign neoplasm and 5 patients with early breast cancer were enrolled and underwent BCS using NIR imaging system. Before surgery, Indocyanine green was injected intratumoraly in benign lesions and peritumoraly for resection margins in early breast cancer. Their pathologic results for resection volume and re-excision rate were compared with those of the patients with 99 benign neoplasm and 203 early breast cancer who were treated with BCS by conventional method, respectively.
Results: In the patients with benign lesions, the mean size in sonography was 1.48±0.98 cm in NIR group and 1.32±0.96cm in conventional group (P=0.4). There was no difference of pathologic lesion size and excised specimen size between these groups (1.57±0.78 cm vs 1.42±0.80 cm in pathologic lesion size; P=0.63, 4.28± 0.48 cm vs 4.15±1.37 cm in specimen size; P=0.73). In NIR group, the ratio of excised specimen/lesion was lesser than that of conventional group (3.19 ±1.40 vs 4.31±3.86; P=0.008). In addition, positive margin rate after BCS in early breast cancer patients was 0% in NIG group and 19.7% in conventional group. And re-excision rate was 0% and 4.4%, respectively.
Conclusions: This study showed that localization using NIR image system could be a feasible method to obtain safe resection margins and optimum resection volumes in patients undergoing breast conserving surgery.
This research was supported by National Cancer Center Grant NCC-1410202-1 & NCC-1410202-2 by the National Cancer Center, Republic of Korea.
Citation Format: Jung S-Y, Lee S, Kim SK. Clinical applications of near infra-red imaging system for localization of non-palpable breast lesions in breast conserving surgery [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-11-10.
Abstract P2-01-19: SentiNot: A way to avoid sentinel node biopsy (SNB) in patients with a preoperative diagnosis of ductal cancer in situ (DCIS)
Authors: Karakatsanis A , Olofsson HM , Eriksson S , Andersson Y , Bergkvist LÅ , Mohammed I , Sundqvist M , Abdsaleh S , Olofsson Bagge R , Sund M , Wärnberg F 
Published: March 2017

Abstract
Background
The risk for node metastasis in preoperative diagnosis of DCIS is low. ASCO guidelines suggest that SNB could be performed when mastectomy is planned or in those cases where the probability of upgrading to invasive cancer postoperatively is high. Despite that, SNB is performed almost in 54% of DCIS procedures in the Uppsala-Örebro Region. Thus, morbidity and resources have to be balanced against the risk of a reoperation.
Methods
Patients with a preoperative diagnosis of DCIS grade 3, grade 2 ≥20mm or planned for a mastectomy will be included. Sienna+ is injected in the breast at the first operation. If the specimen contains invasive cancer, a SNB is performed in another session. Sienna+ can be detected in the axilla at least four weeks after the injection. Endpoints of the trial are the feasibility of detection of the SN as well as how many SNBs are avoided.
Results
In the first 34 cases, six had invasive breast cancer. On reoperation (median 29 days, iqr 10) SNB was successful in four cases with SPIO, one with Tc99 and three cases with blue dye. Totally, the combination of SPIO and blue dye was successful in all cases. No metastases were found.
Patient characteristics Total cohort (N=34)Group with IBC (n=6)Age (yrs)58.6 (54.9, 62.9)60.2 (47.4, 72.9)Size (mm)40.0 (31.2, 62.9)57.7 (27.2, 88.1)DCIS grade29 (26.5%)2 (33.3%) 323 (67.6%)4 (66.7%) na2 (5.9%)0PalpableYes30 (88.2%)1 (16.7%) No4 (11.8%)5 (83.3%)BCSYes22 (64.7%)3 (50%) No12 (35.3%)3 (50%)Transcutaneous counts at the end of the operation266 (196,336)300 (126,474)
Conclusion
It is possible to load the SN with SPIO and reoperate within 4 weeks from the first operation. A rough 83,4% of all patients with DCIS who would have undergone SNB avoided it. Our preliminary results show that 5,6 SNB was avoided for each patient that was operated. SN was detected in all cases with a combination of SPIO and blue dye, but only in 66,7% SPIO. It seems that increased experience with SPIO will increase the effectivity of this procedure.
Citation Format: Karakatsanis A, Olofsson HM, Eriksson S, Andersson Y, Bergkvist LÅ, Mohammed I, Sundqvist M, Abdsaleh S, Olofsson Bagge R, Sund M, Wärnberg F. SentiNot: A way to avoid sentinel node biopsy (SNB) in patients with a preoperative diagnosis of ductal cancer in situ (DCIS) [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-01-19.
Abstract P2-01-23: Long-term follow-up of persistent breast dermopigmentation after sentinel lymph node identification using superparamagnetic iron oxide particles (SIENNA+®)
Authors: Hannebicque K , Boulanger L , Bogart E , Giard S , Chauvet MP , Houpeau JL 
Published: March 2017

Abstract
Background
The French Sentimag study evaluated a non-invasive method for the localization of breast cancer sentinel lymph nodes (SLN) using SIENNA+®, a superparamagnetic iron oxide particles (SPIO), in addition to conventional techniques (radiotracer and blue dye). SIENNA+® was injected subcutaneously into the breast and detected by the SENTIMAG® handheld magnetometer probe. The results showed a good SLN identification performance but a skin discoloration was noted during this study after the SIENNA+® injection. This aim of this study was to assess the long-term duration and appearance of this dermopigmentation.
Methods:
56 patients had participated in Sentimag study in our Center, 6 patients who had undergone mastectomy were excluded. We selected 50 patients who had undergone breast conservative surgery. For these patients, SLN localization was performed by both the conventional method (radiotracer and /or blue dye) and magnetic tracer, SIENNA+®. 47 patients were reviewed retrospectively from January 2015 to April 2015, 1.5 to 2 years after surgery and were assessed for skin discoloration.
Results :
Of the 47 patients, a dermopigmentation, from grade 1 (light yellowing) to grade 3 (dark browning) remained visible at the site of injection of SIENNA+® after 20.2 months [14.4-25.9] in 36.1% of the patients (17/47). 6.4% of 47 patients seen had grade 3 skin discoloration and 29.7% had grade 1 or 2 skin discoloration. Interestingly, no patients reported that persistent staining was a cosmetic or psychological problem.
Conclusions :
The use of SIENNA+® appears as an alternative method to radioisotopes for SLN identification in early breast cancer, but it may result in a prolonged-dermopigmentation at the injection site. To avoid dermopigmentation, it would be interesting to compare different techniques of SIENNA+® injection into the breast (intra-tumoral injection or a deeper periareolar injection) through a randomized trial.
Citation Format: Hannebicque K, Boulanger L, Bogart E, Giard S, Chauvet MP, Houpeau JL. Long-term follow-up of persistent breast dermopigmentation after sentinel lymph node identification using superparamagnetic iron oxide particles (SIENNA+®) [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-01-23.