Early Stage Research Articles

[Transscleral ciliary surgery in glaucoma:from destructive surgery to minimally invasive surgery].

Over the years, there has been significant advancement in the safety and effectiveness of external cyclosurgery for glaucoma. This progress ranges from the initial "cyclodestructive surgery" to modern cycloplasty techniques, expanding beyond end-stage glaucoma treatment. Notably, innovative approaches like micropulse transscleral cycloplasty and ultrasonic cycloplasty are now being employed in earlier stages of glaucoma with better visual acuity, qualifying as minimally invasive procedures. Through a comprehensive review of the historical evolution of external cyclosurgery, elucidation of the mechanisms, clinical outcomes, and potential complications associated with novel cycloplasty techniques, and integration of practical clinical insights, this article aims to furnish clinicians with a profound comprehension of external cyclosurgery for glaucoma.

3D microscaffolds with triple-marker sensitive nanoprobes for studying fatty liver disease in vitro.

Non-alcoholic fatty liver disease (NAFLD) is a heterogeneous condition that encompasses a wide range of liver diseases that progresses from simple hepatic steatosis to the life-threatening state of cirrhosis. However, due to the heterogeneity of this disease, comprehensive analysis of several physicochemical and biological factors that drive its progression is necessary. Therefore, an in vitro platform is required that would enable real-time monitoring of these changes to better understand the progression of these diseases. The earliest stage of NAFLD, i.e. hepatic steatosis, is characterised by triglyceride accumulation in the form of lipid vacuoles in the cytosol of hepatocytes. This fatty acid accumulation is usually accompanied by hepatic inflammation, leading to tissue acidification and dysregulated expression of certain proteases such as matrix metalloproteinases (MMPs). Taking cues from the biological parameters of the disease, we report here a 3D in vitro GelMA/alginate microscaffold platform encapsulating a triple-marker (pH, MMP-3 and MMP-9) sensitive fluorescent nanoprobe for monitoring, and hence, distinguishing the fatty liver disease (hepatic steatosis) from healthy livers on the basis of pH change and MMP expression. The nanoprobe consists of a carbon nanoparticle (CNP) core, which exhibits intrinsic pH-dependent fluorescence properties, decorated either with an MMP-3 (NpMMP3) or MMP-9 (NpMMP9) sensitive peptide substrate. These peptide substrates are flanked with a fluorophore-quencher pair that separates on enzymatic cleavage, resulting in fluorescence emission. The cocktail of these nanoprobes generated multiple fluorescence signals corresponding to slightly acidic pH (blue) and overexpression of MMP-3 (green) and MMP-9 (red) enzymes in a 3D in vitro fatty liver model, whereas no/negligible fluorescence signals were observed in a healthy liver model. Moreover, this platform enabled us to mimic fatty liver disease in a more realistic manner. Therefore, this 3D in vitro platform encapsulating triple-marker sensitive fluorescent nanoprobes would facilitate the monitoring of the changes in pH and MMP expression, thereby enabling us to distinguish a healthy liver from a diseased liver and to study liver disease stages on the basis of these markers.

Gonadotropin elevation is ootoxic to ovulatory oocytes and inhibits oocyte maturation, and activin decoy receptor ActRIIB:Fc therapeutically restores maturation

BackgroundElevated FSH often occurs in women of advanced maternal age (AMA, age ≥ 35) and in infertility patients undergoing controlled ovarian stimulation (COS). There is controversy on whether high endogenous FSH contributes to infertility and whether high exogenous FSH adversely impacts patient pregnancy rates.MethodsThe senescence-accelerated mouse-prone-8 (SAMP8) model of female reproductive aging was employed to assess the separate impacts of age and high FSH activity on the percentages (%) of viable and mature ovulated oocytes recovered after gonadotropin treatment. Young and midlife mice were treated with the FSH analog equine chorionic gonadotropin (eCG) to model both endogenous FSH elevation and exogenous FSH elevation. Previously we showed the activin inhibitor ActRIIB:Fc increases oocyte quality by preventing chromosome and spindle misalignments. Therefore, ActRIIB:Fc treatment was performed in an effort to increase % oocyte viability and % oocyte maturation.ResultsThe high FSH activity of eCG is ootoxic to ovulatory oocytes, with greater decreases in % viable oocytes in midlife than young mice. High FSH activity of eCG potently inhibits oocyte maturation, decreasing the % of mature oocytes to similar degrees in young and midlife mice. ActRIIB:Fc treatment does not prevent eCG ootoxicity, but it restores most oocyte maturation impeded by eCG.ConclusionsFSH ootoxicity to ovulatory oocytes and FSH maturation inhibition pose a paradox given the well-known pro-growth and pro-maturation activities of FSH in the earlier stages of oocyte growth. We propose the FOOT Hypothesis (“FSH OoToxicity Hypothesis), that FSH ootoxicity to ovulatory oocytes comprises a new driver of infertility and low pregnancy success rates in DOR women attempting spontaneous pregnancy and in COS/IUI patients, especially AMA women. We speculate that endogenous FSH elevation also contributes to reduced fecundity in these DOR and COS/IUI patients.Restoration of oocyte maturation by ActRIB:Fc suggests that activin suppresses oocyte maturation in vivo. This contrasts with prior studies showing activin A promotes oocyte maturation in vitro. Improved oocyte maturation with agents that decrease endogenous activin activity with high specificity may have therapeutic benefit for COS/IVF patients, COS/IUI patients, and DOR patients attempting spontaneous pregnancies.

Cutaneous biomarkers of therapeutic efficacy in early treatment of hereditary ATTR amyloid polyneuropathy with tafamidis.

ATTR (ATTRv) amyloidosis neuropathy is characterized by progressive sensorimotor and autonomic nerve degeneration secondary to amyloid deposition caused by a misfolded transthyretin protein (TTR). Small nerve fiber neuropathy is an early clinical manifestation of this disease resulting from the dysfunction of the Aδ and C small nerve fibers. Tafamidis, a selective TTR stabilizer, has proven its efficacy in the earlier stages of hATTR. To evaluate the clinical course and utility of cutaneous pathological biomarkers in patients with ATTR amyloidosis treated with tafamidis compared to control patients. Forty patients diagnosed with early stages of ATTRv amyloidosis (polyneuropathy disability [PND] scores 0-II) underwent small and large nerve fiber neurological evaluations, and annual skin biopsies for intraepidermal nerve fiber density (IENFD) and amyloid deposition index (ADI) estimation. Thirty patients were allocated to receive tafamidis, and 10 patients served as controls. Tafamidis pharmacokinetics analysis was performed in patients who received the treatment. At baseline, 12% of patients in stage PND 0 and 28% in PND I displayed small nerve fiber denervation in the distal thigh, whereas 23% and 38%, respectively, in the distal leg. Similarly, 72% and 84% had amyloid deposition in the distal thigh and 56% and 69% in the distal leg. Following 1 year of treatment, the tafamidis group showed significant clinical improvement compared to the control group, revealed by the following mean differences (1) -9.3 versus -4 points (p = <.00) in the patient's neuropathy total symptom score 6 (NTSS-6) questionnaire, (2) -2.5 versus +2.8 points (p = <.00) in the Utah Early Neuropathy Score (UENS), and (3) +1.2°C versus -0.6 (p = .01) in cold detection thresholds. Among the patients who received tafamidis, 65% had stable or increased IENFD in their distal thigh and 27% in the distal leg. In contrast, all patients in the control group underwent denervation. The ADI either decreased or remained constant in 31% of the biopsies in the distal thigh and in 24% of the biopsies in the distal leg of the tafamidis-treated patients, whereas it rose across all the biopsies in the control group. At the 4-year follow-up, the tafamidis group continued to display less denervation in the distal thigh (mean difference [MD] of -3.0 vs. -9.3 fibers/mm) and the distal leg (mean difference [MD] -4.9 vs. -8.6 fibers/mm). ADI in tafamidis-treated patients was also lower in the distal thigh (10 vs. 30 amyloid/mm2) and the distal leg (23 vs. 40 amyloid/mm2) compared to control patients. Plasma tafamidis concentrations were higher in patients with IENFD improvement and in patients with reduced amyloid deposition. Patients without amyloid deposition in the distal leg at baseline displayed delayed disease progression at 4 years. Cutaneous IENFD and amyloid deposition assessments in the skin of the distal thigh and distal leg are valuable biomarkers for early diagnosis of ATTR amyloidosis and for measuring the progression of small nerve fiber neuropathy. Early treatment with tafamidis slows the clinical progression of the disease, skin denervation, and amyloid deposition in the skin. Higher plasma concentrations of tafamidis are associated with better disease outcomes, suggesting that increasing the drug dose could achieve better plasma concentrations and response rates. This study describes the longest small nerve fiber neuropathy therapeutic trial with tafamidis and is the first to report small fiber symptoms, function, and structural assessments as outcomes.

Integrating artificial intelligence techniques for advancements in colorectal cancer management: navigating past and predicting future direction.

Artificial Intelligence (AI) in the last few years has emerged as a valuable tool in managing colorectal cancer, revolutionizing its management at different stages. In early detection and diagnosis, AI leverages its prowess in imaging analysis, scrutinizing CT scans, MRI, and colonoscopy views to identify polyps and tumors. This ability enables timely and accurate diagnoses, initiating treatment at earlier stages. AI has helped in personalized treatment planning because of its ability to integrate diverse patient data, including tumor characteristics, medical history, and genetic information. Integrating AI into clinical decision support systems guarantees evidence-based treatment strategy suggestions in multidisciplinary clinical settings, thus improving patient outcomes. This narrative review explores the multifaceted role of AI, spanning early detection of colorectal cancer, personalized treatment planning, polyp detection, lymph node evaluation, cancer staging, robotic colorectal surgery, and training of colorectal surgeons.

Survival benefit of radical prostatectomy in patients with advanced TURP-diagnosed prostate cancer: a population-based real-world study

ObjectivesA considerable number of patients are diagnosed with prostate cancer (PCa) by transurethral resection of the prostate (TURP). We aimed to evaluate whether radical prostatectomy (RP) brings survival benefits for these patients, especially in the elderly with advanced PCa.Patients and methodsWe used the Surveillance, Epidemiology, and End Results (SEER) database to obtain PCa cases diagnosed with TURP. After the propensity matching score (PSM) for case matching, univariate, multivariate, and subgroup analyses were performed to investigate whether RP impacts the survival benefit.Results4,677 cases diagnosed with PCa by TURP from 2010 to 2019 were obtained, including 1,313 RP patients and 3,364 patients with no RP (nRP). 9.6% of RP patients had advanced PCa. With or without PSM, cancer-specific mortality (CSM) and overall mortality (OM) were significantly reduced in the RP patients compared to the nRP patients, even for older (> 75 ys.) patients with advanced stages (all p < 0.05). Except for RP, younger age (≤ 75 ys.), being married, and earlier stage (localized) contributed to a significant reduction of CSM risk (all p < 0.05). These survival benefits had no significant differences among patients of different ages, married or single, and at different stages (all p for interaction > 0.05).ConclusionsBased on this retrospective population-matched study, we first found that in patients diagnosed with PCa by TURP, RP treatment may lead to a survival benefit, especially a reduction in CSM, even in old aged patients (> 75 ys.) with advanced PCa.

The origin of betaine in mouse oocytes and preimplantation embryos†.

Betaine has important roles in preimplantation mouse embryos, including as an organic osmolyte that functions in cell volume regulation in the early preimplantation stages and as a donor to the methyl pool in blastocysts. The origin of betaine in oocytes and embryos was largely unknown. Here, we found that betaine was present from the earliest stage of growing oocytes. Neither growing oocytes nor early preantral follicles could take up betaine, but antral follicles were able to transport betaine and supply the enclosed oocyte. Betaine is synthesized by choline dehydrogenase, and female mice lacking Chdh did not have detectable betaine in their oocytes or early embryos. Supplementing betaine in their drinking water restored betaine in the oocyte only when supplied during the final stages of antral follicle development but not earlier in folliculogenesis. Together with the transport results, this implies that betaine can only be exogenously supplied during the final stages of oocyte growth. Previous work showed that the amount of betaine in the oocyte increases sharply during meiotic maturation due to upregulated activity of choline dehydrogenase within the oocyte. This betaine present in mature eggs was retained after fertilization until the morula stage. There was no apparent role for betaine uptake via the SIT1 (SLC6A20) betaine transporter that is active at the 1- and 2-cell stages. Instead, betaine was apparently retained because its major route of efflux, the volume-sensitive organic osmolyte - anion channel, remained inactive, even though it is expressed and capable of being activated by a cell volume increase.

Primordial germ cell DNA demethylation and development require DNA translesion synthesis

Mutations in DNA damage response (DDR) factors are associated with human infertility, which affects up to 15% of the population. The DDR is required during germ cell development and meiosis. One pathway implicated in human fertility is DNA translesion synthesis (TLS), which allows replication impediments to be bypassed. We find that TLS is essential for pre-meiotic germ cell development in the embryo. Loss of the central TLS component, REV1, significantly inhibits the induction of human PGC-like cells (hPGCLCs). This is recapitulated in mice, where deficiencies in TLS initiation (Rev1-/- or PcnaK164R/K164R) or extension (Rev7 -/-) result in a > 150-fold reduction in the number of primordial germ cells (PGCs) and complete sterility. In contrast, the absence of TLS does not impact the growth, function, or homeostasis of somatic tissues. Surprisingly, we find a complete failure in both activation of the germ cell transcriptional program and in DNA demethylation, a critical step in germline epigenetic reprogramming. Our findings show that for normal fertility, DNA repair is required not only for meiotic recombination but for progression through the earliest stages of germ cell development in mammals.

Radio Emission from High-redshift Active Galactic Nuclei in the JADES and CEERS Surveys

Recent calculations indicate that radio emission from quasars at z ∼ 6–7 could be detected at much earlier stages of evolution, at z ∼ 14–15, by the Next-Generation Very Large Array (ngVLA) and the Square Kilometer Array (SKA). However, the James Webb Space Telescope has now discovered less luminous active galactic nuclei (AGNs) at z > 4 and a few massive black holes (BHs) at z > 10, which may be the progenitors of supermassive black holes (SMBHs) but at different stages of growth. Radio detections of these new AGNs would provide complementary measures of their properties and those of their host galaxies. Here we estimate radio flux densities for 19 new AGNs found by the JADES, CEERS, and UNCOVER surveys. We find that ngVLA should be able to detect most of these sources in targeted surveys with integration times of 10–100 hr (and in just 1 hr for a few of them) but most would require at least 100 hr of SKA time in spite of its greater sensitivities at low frequencies. In some cases, radio emission from the BH can be distinguished from that of H ii regions and supernovae in their host galaxies, which could be used to estimate their star formation rates. Such detections would be yet another example of the useful synergies between near-infrared and radio telescopes in SMBH science in the coming decade.

Abstract PO4-10-03: Relationship Between Insurance Status and Breast Cancer Care in Young Mexican Women

Abstract BACKGROUND In Mexico, 71% of the population is covered by public health insurance, while 2% are privately insured and 27% are uninsured. Previous studies have demonstrated differences in breast cancer (BC) care according to type of healthcare coverage in the United States and Latin America. Most studies have observed that privately insured patients are diagnosed at earlier stages, have less symptom-to-diagnosis and treatment delays, and have better overall outcomes. The aim of this analysis was to evaluate whether differences in BC care exist among young women with breast cancer (YWBC), ≤40 years at diagnosis, depending on their insurance status in Mexico. METHODS Data was collected from the Mexican prospective cohort of YWBC Joven &amp; Fuerte. Patient surveys and researcher follow-ups were used to collect data related to clinicopathologic features, time from symptom onset to seeking medical assistance and treatment, and perceived quality of information received and medical staff support. Results were analyzed using SPSS. RESULTS Information about health insurance status was available for 608 patients: 565 (93%) had public coverage and 43 (7%) had private coverage. Table 1 describes the clinicopathologic characteristics of both groups. Patients with private insurance were diagnosed at earlier stages, 0-II vs. III-IV (81% vs. 16%), than those with public insurance (50% vs. 45%) (p&amp;lt; 0.001). No difference was found on the distribution of BC subtypes among both groups. A total of 573 (94%) patients had registered information regarding the time from symptom onset to contacting a medical professional. Only 18 (3%) were diagnosed with imaging studies (5% and 3% with private and public coverage, respectively). More privately insured patients (95%) contacted a healthcare professional during the first 3 months since symptom onset than publicly insured patients (74%) (p=0.003). Information regarding the time from contacting a healthcare professional to treatment start was available for 472 (78%) patients. There was no difference in the proportion of patients who started treatment ≤3 months after arriving with a healthcare professional according to type of health insurance (76% private vs. 67% public coverage; p=0.2). The majority of patients (96%) considered having adequate support from the medical staff (100% private vs. 94% public). Compared to those with public coverage, more privately insured patients perceived receiving adequate information about their treatment (79% private vs. 50% public; p&amp;lt; 0.001). There were no differences between groups regarding the perceived quality of information regarding their diagnosis (p=0.06) and continuous BC care (p=0.6). CONCLUSION Patients with private coverage were diagnosed at earlier stages, reported contacting a healthcare professional earlier, and perceived receiving adequate information regarding their treatment more often than patients with public coverage. These results coincide with previously reported differences between patients with private and public health insurance. Interestingly, there were no differences between the delays from the first visit to treatment initiation and perceived support from the medical staff depending on type of healthcare coverage. Given the observed differences between privately and publicly insured patients, population-based interventions to increase awareness of alarm signs for BC and widespread access to quality patient education resources could help decrease the disparities between these groups. Table 1. Clinicopathologic features of BC depending on insurance status Citation Format: David G Gonzalez-Sanchez, Arantxa Ramirez-Cisneros, Alejandra Platas, Alejandra Platas, Daniela Vazquez-Juarez, Fernanda Mesa-Chavez, Ana Ferrigno, Alejandro Mohar, Alan Fonseca, Cynthia Villarreal-Garza. Relationship Between Insurance Status and Breast Cancer Care in Young Mexican Women [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-10-03.

Abstract PO4-15-01: A multi-center prospective cohort study to evaluate the presence of circulating tumor cells using the Epic Sciences platform among women with metastatic breast cancer

Abstract Background: The presence of tumor cells (or their components) in the blood of women with a history of early breast cancer has the potential to herald the development of metastatic recurrence at its earliest stages. Such early detection could potentially lead to novel prevention strategies, but it requires a sensitive assay. Objective: To use the Epic Sciences platform to detect and enumerate CTCs in blood samples from patients with an established diagnosis of metastatic breast cancer (MBC), prior to initiation of 1st line systemic therapy in the metastatic setting. Methods: We conducted a multi-center prospective cohort study to evaluate the presence of CTCs using the Epic Sciences platform among patients with a new diagnosis of MBC. Men or women age 18 to 85 were included, irrespective of breast cancer subtype. Patients with a prior or concurrent malignancy whose natural history or treatment had the potential to interfere with the detection of MBC in a liquid biopsy were excluded. A one-time blood draw was performed before patients received any local or systemic therapy in the metastatic setting. In addition, those with recurrent disease must have been off any systemic adjuvant therapy for ≥3 weeks prior to blood collection. Two 5 mL blood samples were obtained for CTC identification and enumeration. CTC identification was based on immunofluorescence analysis using Epic Sciences platform as previously described (Ueno et al 2017). The presence of CTCs was correlated with clinical and pathological features, which were abstracted from medical records and pathology reports. The association between the presence of CTCs and clinical/pathologic characteristics was tested using Fisher’s exact test for categorical variables and t-test or Wilcoxon rank sum tests for numerical variables. All analyses were performed using the R software package. Results: 100 patients were recruited between February 2021 and January 2023 at five academic oncology centres in Ontario and Quebec, Canada. 95 patients had evaluable blood for analysis and 5 did not due to blood age and/or insufficient blood volume. Six patients were excluded after providing a blood sample because tissue biopsy ultimately revealed a 2nd primary tumor (n=4) or benign tissue (n=2). Hence, 89 patients with a clinical diagnosis of MBC and with evaluable blood for CTC analyses were ultimately included in our cohort. The average age of patients was 61 years. Most patients (n=49, 55%) had a prior history of early breast cancer, 38 (43%) had de-novo metastatic disease and prior breast cancer history was unknown for 2 patients. 50 (66%) of patients had visceral metastatic disease. The most common sites of metastases included bone (62%), lung (30%), liver (29%) and lymph nodes (17%). 63 of 89 patients (71%) had detectable CTCs at baseline, prior to any local or systemic treatment in the metastatic setting. The median number of detectable CTCs per 5mL sample was 2 (IQR 8.5) and the range was 0 – 12,798. Twenty nine of 89 (33%) patients had 5 or more CTCs detected per 5ml blood. The proportion of patients with detectable CTCs was numerically highest (n=39/51, 76%) among patients with hormone receptor (HR)+/HER2-ve breast cancer, followed by HER2+ (n=16/22, 73%) and triple negative (n=8/13, 62%) disease. Associations between CTC detection with prior history of early breast cancer, sites of metastatic disease and disease burden will also be presented. Conclusions: Approximately 3 in 4 women with newly diagnosed metastatic breast cancer have detectable CTCs using the Epic Sciences platform prior to initiation of first line systemic therapy. CTCs may be a promising tool for the monitoring of breast cancer recurrence and will be investigated in an ongoing Canadian prospective observational study that aims to elucidate biomarkers of late breast cancer recurrence. Citation Format: Katarzyna Jerzak, Pamela Goodwin, Marguerite Ennis, Christine Brezden-Masley, Nathaniel Bouganim, Mark Basik, Arushi Jain, Giuseppe Di Caro, Rick Wenstrup, Nadine Hartmann, Megan Slade, Ana Elisa Lohmann. A multi-center prospective cohort study to evaluate the presence of circulating tumor cells using the Epic Sciences platform among women with metastatic breast cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-15-01.

Abstract PO5-05-07: Efficacy and safety of first line CDK4/6i plus endocrine therapy for patients with HR+/HER2- metastatic breast cancer:initial real-world experience at Ho Chi Minh city oncology hospital, Viet Nam

Abstract Background The cyclin-dependent kinases (CDK)4/6 inhibitors (CDK4/6i) palbociclib and ribociclib have been available in Viet Nam since 2021 for the treatment of hormone receptor-positive metastatic breast cancer (MBC). Data regarding CDK4/6i benefit in Vietnamese MBC are sparse. This study evaluated the impact of CDK4/6i in first-line HR+/HER2− MBC using real-world experience at Ho Chi Minh City Oncology Hospital. Methods A single-institution study analyzed retrospective data from 108 patients treated with palbiciclib or ribociclib with endocrine therapy in first-line in HR+/HER2– MBC patients at Ho Chi Minh city oncology hospital. Patient characteristics, PFS, treatment response, and toxicity profiles were analyzed. Results Baseline patients’ characteristics are detailed in Table 1.The median follow-up was 12.5 months (95% CI: 10.9-13.3). Best responses by the line of therapy are in Table 2. The median PFS has still not been reached. The PFS rates at 6, 12 and 18 months were 94.4%, 93.5% and 91.5%, respectively. Patients with bone-only metastasis have a better PFS than visceral metastasis (p = 0.059). Patients with recurrent disease had shorter PFS (p = 0.083) than those presenting with de novo metastasis. The most common reasons for toxicity were neutropenia (92.6%), anemia (35.2%), thrombopenia (22.2%) and the incidence of diarrhea was only 2,8%. None were related to QTc-prolongation Conclusion The real-world effectiveness and safety with CDK4/6 inhibitors in our institution HR+/HER2– MBC patients mimics that observed in the clinical trials. This finding supports the use of CDK4/6i in combination with endocrine therapy as standard of care for HR+/HER2- MBC in Viet Nam. The study also suggests the need for more study of under-represented minority populations. Further studies are ongoing to validate these findings incorporating additional cancer centers. Keywords: advanced/metastatic breast cancer, Asia, CDK4/6 inhibitors, real world Table 1. Patient Demographics and Baseline Disease Characteristics (N=108) Characteristics N (%) Age, y: Median 52 Stage Recurrent from earlier stage, stages 0–III 67 (62) De novo, newly diagnosed stage IV at enrolment 41 (38) DFI &amp;lt; 12 months 5 (4.6) 12-24 months 12 (11.1) &amp;gt; 24 months 51 (47.2) No. of metastatic sites 1 49 (45.4) 2 37 (34.3) ≥ 3 22 (20.4) Metastasis pattern Bone only 30 (27.8) Other 78 (72.2) CDK4/6i Palbociclib 35 (32.4) Ribociclib 73 (67.6) Partner ET AIs 94 (87) Fulvestrant 14 (13) Table 2. Best responses of therapy (N=108) Best responses (N,%) CR 2 (1.9) PR 29 (26.9) SD 67 (62.1) PD 10 (9.3) Total 108 (100) ORR 31 (28.8) CBR 76 (70.3) Table. Citation Format: Hoang-Quy Nguyen, Hong-Duc Phan-Thi, Thi-Hong-Van Le. Efficacy and safety of first line CDK4/6i plus endocrine therapy for patients with HR+/HER2- metastatic breast cancer:initial real-world experience at Ho Chi Minh city oncology hospital, Viet Nam [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-05-07.

Abstract PO4-03-03: Conditional Use of Serial Serum Tumor Markers CA 15.3 and CEA as Predictors of Disease Relapse in Patients with High-Risk Early-Stage Breast Cancer

Abstract Background: Because of the diagnosis at an earlier stage, and the wider availability of more effective therapies, treatment outcomes of patients with breast cancer are getting much better resulting in an increasing proportion of patients surviving their disease, however, a significant percentage of patients with early-stage disease at initial diagnosis may eventually relapse. Residual tumor cells can remain dormant for many years before causing tumor recurrence. Physical examination and mammography are strongly recommended in surveillance guidelines, but data on routine blood tests or imaging is lacking. The revolution made in recent years, following the approval of novel targeted and immunotherapy agents, may have better outcome if started earlier in the disease course with the lowest tumor burden and in patients with good performance status and adequate organs’ function. Additionally, early detection of oligometastatic disease can give a chance for cure for a subset of these patients. Levels of CA15.3 and CEA are used in decision making in the metastatic setting but their role in surveillance is still controversial. We aim to investigate whether serial measurement of tumor markers, when it correlates with tumor bulk (conditional), can detect early asymptomatic recurrence among subset of patients with high-risk early-stage breast cancer. Methods: Patients with high-risk early-stage breast cancer were invited to participate. High-risk features include any of the following: large tumor size (T3/4), grade-3, node-positive, triple-negative, HER2-positive or hormone receptor (HR)-negative disease. Patients were considered eligible if they have elevated CA15.3 and/or CEA at baseline, that subsequently normalize 6 weeks after surgical resection; upfront or following neoadjuvant chemotherapy. Serial testing of the elevated CA15.3 and/or CEA is done every 2 months thereafter, samples are cryopreserved and will not be processed until disease relapse. As such results will not be used for clinical decisions. The study is still ongoing, we here present an interim analysis evaluating the correlation between certain tumor characteristics and elevated serum marker(s) levels. Results: Since the launch of the study, 367 patients deemed high-risk by the above criteria, accepted to participate. Abnormal baseline CA15.3 and/or CEA was found in 110 (30%); 65 (18%) patients had elevated CA 15.3 while elevated CEA was found in 45 (12%) patients. Abnormal CA 15.3 was found in 27% of cases with T3/4 disease compared with 13% in T1/2 disease (p=0.001), and in 21% of node positive disease versus 7% in node negative disease (p=0.002), in 9% of HR negative disease versus 20% in HR-positive (P=0.038). Tumor grade and HER2 status were not associated with significant difference. Regarding CEA, abnormal levels were detected in 24% of HER2-positive disease compared to 5% of HER2-negative disease (P&amp;lt; 0.0001). Other tumor features were not associated with significant difference. Conclusions: Only a third of patients with high-risk early-stage breast cancer had high level of serum tumor markers at initial diagnosis and thus such markers may correlate with tumor presence. Large tumors (T3/4) and node positive disease were associated with high CA15.3 but not CEA. HER2-stutus and tumor grade had no effect on CA15.3 level, while HER2 positive disease correlated well with high CEA. Citation Format: Sarah Edaily, Baha' Sharaf, Maher Sughayer, Lina Yousef, Hala Abu-Fares, Sarah Abdel-Razeq, Hala Abu-Jaish, Mahmoud Abunasser, Suhaib Khater, Anas Zayed, Osama El Khatib, Rahaf Rahal, Hazem Abdulelah, Maria Shraim, Hikmat Abdel-Razeq. Conditional Use of Serial Serum Tumor Markers CA 15.3 and CEA as Predictors of Disease Relapse in Patients with High-Risk Early-Stage Breast Cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-03-03.

Abstract PO2-26-01: Prognostic Role of HER2 Expression in Patients with ER-positive/HER2-negative Breast Cancer. Results from a Population-Based Cancer Registry Study

Abstract Background: Estrogen Receptor (ER)-positive (+)/Human Epidermal Growth Factor Receptor 2 (HER2)-negative (-) breast cancers (BCs) express variable protein levels of HER2, which can influence prognosis. Materials and Methods: This cohort study was conducted using data on all consecutive patients (pts) diagnosed with BC between 2005 and 2017, which were systematically and prospectively collected by the Emilia-Romagna Cancer Registry (Provinces of Piacenza, Parma, Modena, and Ferrara), Italy. The study included 13527 pts with ER+/HER2- BC. Tumors were classified by HER2 IHC score (0 [n=7155], 1+ [n=5186], or 2+ with negative FISH [n=1186]). Comparisons of clinicopathologic characteristics and disease outcome were performed. Results: BCs with late-stage diagnosis, high histological grade, or high proliferative rate were more likely to be HER2 1+ or 2+/FISH- in comparison with earlier stage, low-grade, or low-proliferative tumors (lower bounds of 95% confidence intervals [CIs] for odds ratios [ORs] &amp;gt; 1). BCs with high expression of ER (≥ 80% [n=12383]) were more enriched with HER2 1+ (OR 1.39; 95%CI, 1.2-1.6) or 2+/FISH- tumors (OR 1.26; 95%CI, 1.0-1.6) than ER-low/moderate (1-79% [n=1144]) ones. The 5-year overall survival (OS) for HER2 1+ BCs was lower than that for HER2 0 or 2+/FISH- tumors (P = 0.0018). This finding was confirmed also after stratification by ER status (low/moderate and high expression; P = 0.07 and 0.007, respectively). By multivariate logistic regression, the following variables were significantly associated with HER2 1+, or 2+/FISH- expression compared to HER2 0 tumors: age at diagnosis &amp;lt; 50, high histological grade, ER expression ≥ 80% (lower bounds of 95% CIs for ORs &amp;gt; 1). Multivariate Cox's regression analysis showed that histological grades G2 and G3 were independently associated with poor survival vs. G1 tumors (HR 1.2; 95%CI, 1.1-1.3 and 1.3; 95%CI, 1.1-1.4, respectively). Furthermore, HER2 2+/FISH- status was significantly associated with better survival in comparison with HER2 0 tumors (HR 0.82; 95%CI, 0.7-0.9). This finding was confirmed also after stratification by ER status (ER 1-79% and ≥ 80%). No significant difference in OS was observed between HER2 1+ and 0 BCs. Conclusions: The putative worse prognostic impact of HER2 expression in pts with ER-positive/HER2-negative BCs was not confirmed. The better outcome observed in pts with HER2 2+/FISH- BCs may be related to the known FISH-negative (HER2-non-amplified) status of this subgroup. These findings may help identify optimal patient inclusion criteria for clinical trials with novel anti-HER2 therapies in ER-positive/HER2-negative disease. Citation Format: Antonino Musolino, Olga Serra, Benedetta Pellegrino, Chiara Tommasi, Daniele Zanoni, Fabio Faccini, Maria Michiara, Paolo Sgargi, Giuliano Carrozzi, Stefano Ferretti, Veruska Grossi, Alessandra Ravaioli, Federica Zamagni, Fabio Falcini. Prognostic Role of HER2 Expression in Patients with ER-positive/HER2-negative Breast Cancer. Results from a Population-Based Cancer Registry Study [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-26-01.

Abstract PO5-04-07: Characteristics of patients with metastatic HR+/HER2+ breast cancer in a Kaiser Permanente observational multi-site cohort

Abstract Background: Approximately 10% of female breast cancer and 13% of male breast cancer tumors express hormone-receptor positive/human epidermal growth factor receptor 2 positive (HR+/HER2+) molecular subtype. Since HER2+ targeted agents were made available nearly 10 years ago, there has been limited innovation in treatment of patients with HR+/HER2+ metastatic breast cancer (mBC) to improve clinical outcomes. This study aims to characterize the HR+/HER2+ mBC patient population by describing demographic and clinical characteristics and treatment patterns in a racially diverse, insured, population-based cohort. This study includes data from 4 regions of the Kaiser Permanente (KP) health system – Northwest, Northern California, Southern California, and Colorado. Methods: Eligible study patients were adults age &amp;gt; 18 years with HR+/HER2+ invasive breast cancer who were either initially diagnosed with Stage IV disease or were diagnosed at an earlier stage and experienced recurrence with distant metastasis between 6/8/2012 and 12/31/2019 with data collected through 12/31/2021. Select information was extracted from electronic health records (EHR) of eligible patients and combined with manually abstracted data to create the analytic dataset. Breast cancer recurrence was identified by tumor registrars or obtained through a breast cancer recurrence detection algorithm and verified by chart review. Chart reviewers collected data on metastatic sites, disease progression, reasons for stopping systemic therapy medications, and verified use of medications. Preliminary results: During the study period, we identified 492 subjects with HR+/HER2+ mBC. Seven were men and 485 were women. 221 (45%) had de novo Stage IV breast cancer at diagnosis and 271 (55%) had recurrent mBC. Median age at metastatic diagnosis was 59 years for de novo cases and 62 years for recurrent cases. The cohort was racially and ethnically diverse, with 13% Asian subjects (11% de novo, 15% recurrent), 12% Black subjects (10% de novo, 14% recurrent), and 66% White subjects (71% de novo, 63% recurrent). 18% of cases were recorded as Hispanic (19% de novo, 18% recurrent) in the EHR. Major sites of metastases, including bone, liver, lung, brain, and distant lymph nodes, were similar in both the de novo and recurrent groups. Among women (see table below), 28 (6%) received no systemic treatment for their metastatic disease (9% de novo, 3% recurrent). Among women with systemic treatment, 80% received anti-HER2 agents (90% de novo, 73% recurrent). Of women with recurrent mBC who received systemic therapy, 56% had received anti-HER2 therapy in the adjuvant setting. Conclusion: This population-based study of a racially diverse cohort examined EHR-derived data to describe current demographics and treatment patterns for HR+/HER2+ mBC in an insured patient population. Over 90% of women received systemic therapy, and the majority of those received anti-HER2 therapy per current treatment guidelines. Future analyses will focus on clinical outcomes in this cohort with a goal of informing how prognosis can be improved. Table. Citation Format: Sheila Weinmann, Debra Ritzwoller, Marilyn Kwan, Reina Haque, Stella Stergiopoulos, Ashley Cha, Eric Gauthier, Daniel Sapp, John Dickerson, Kathryn Richert-Boe, Weiming Hu, Nikki Carroll, Lie Hong Chen, Valerie Lee, Ravi Zalavadia, Maureen O'Keeffe-Rosetti. Characteristics of patients with metastatic HR+/HER2+ breast cancer in a Kaiser Permanente observational multi-site cohort [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-04-07.

EXPRESS: Flowers and Bees: Spatial Network Effects in the Adoption of a Sharing-Economy Platform

This paper empirically analyzes the spatiotemporal diffusion of a car-sharing platform that connects consumers with individual car owners. We develop a model of adoption by both consumers and providers that accommodates network effects across locations, as consumers travel to pick up cars, and information asymmetry, as only the providers’ locations are shown on a map. We apply it to data at the earliest stage of the platform and find that proximity and consumer mobility play a significant role in the spatial network effects of existing providers on consumers across locations. Consequently, the impact of additional supply at a destination on consumer adoptions at an origin varies by origin-destination pair and is asymmetric, as certain destinations draw more visitors than others and given differences in local characteristics. In contrast, existing consumers have a limited impact on provider adoptions. Through seeding experiments, we investigate how the geographic distribution of initial participants impacts the platform diffusion and find that targeting the supply side in big cities leads to the highest platform growth. We also use our parameter estimates to measure the long-term impact of a local promotional campaign and show that it is mostly local despite spatial network effects.

Abstract PO2-13-08: Cell-free orphan noncoding RNAs and AI enable early detection of invasive breast cancer and ductal carcinoma in-situ

Abstract Background Earlier detection of breast cancer through mammography screening has reduced disease-specific mortality; however, confounding issues such as technical challenges, breast density, and tumor size can result in false negatives and ultimately later stage diagnosis. Next generation liquid biopsy has the potential to complement mammography and enable earlier detection for more women. We have previously demonstrated high sensitivity and specificity for early detection of invasive breast cancer (IBC) by utilizing a novel category of cancer-associated small RNAs, termed orphan noncoding RNAs (oncRNAs), through a liquid biopsy platform. Here, we further improve the ability to detect breast cancer in a larger, multi-source cohort through an AI-driven approach and demonstrate potential for detection of ductal carcinoma in-situ (DCIS). Methods We utilized The Cancer Genome Atlas (TCGA) small RNA-seq database to discover a library of 20,538 oncRNAs, through a female-specific analysis, that were significantly enriched among 1,103 breast tumors compared to 349 normal tissue samples spanning multiple tissue sites. The diagnostic performance of these oncRNAs were assessed in an independent cohort of archived serum samples from 663 female individuals, sourced from Indivumed (Hamburg, Germany), Proteogenex (Inglewood, CA), and MT Group (Los Angeles, CA), including 279 breast cancer patients of various stages (221 IBC and 58 DCIS; mean age: 57.0 ± 13.8 years; ever-smoker: 25.8%) and 304 age-matched controls (mean age: 58.5 ± 13.9 years; ever-smoker: 23.4%) without breast cancer. All samples were collected between 2010–2022 at time of diagnosis for breast cancer patients. We sequenced the small RNA content of these samples at an average depth of 25.28 ± 9.37 million 50-bp single-end reads. We detected 18,025 (87.8%) unique breast cancer-specific oncRNA species within at least one sample from the study cohort. We then trained a generative AI model using 5-fold cross-validation to predict cancer status for all samples. Results Our oncRNA-based model achieved an overall AUC of 0.95 (95% CI, 0.93–0.97) for prediction of IBC versus cancer-free controls with a sensitivity of 0.87 (0.82–0.91) at 90% specificity. We observed high sensitivities, also at 90% specificity, across all tumor stages and tumor sizes (Table 1). Sensitivities for the earliest stage and smallest tumor size were 0.87 (0.78–0.93) and 0.81 (0.61–0.93) for Stage I (n=83) and T1a–b ( &amp;gt;1mm to ≤10mm; n=26), respectively. Additionally, in a small single-source cohort, we also saw high model accuracy and sensitivity for DCIS, which we aim to confirm in additional cohorts. While our overall cancer cohort primarily consisted of individuals with luminal breast cancer, our model had high sensitivities across all breast cancer subtypes at 0.90 (0.84–0.94), 0.73 (0.59–0.85), and 0.86 (0.42–1.0) for luminal (n=181), HER2 positive (n=49), and triple negative (n=7), respectively. Conclusions We further demonstrate the potential utility of oncRNAs as a blood-based biomarker using an AI algorithm for sensitive and accurate early detection of breast cancer in a large cohort. Additionally, we have shown that this oncRNA-based assay performs well in detecting small, early-stage invasive breast tumors, with potential to detect precursors of breast cancer. Table 1: Model sensitivity in breast cancer by tumor stage and size For each tumor stage and size, as defined by the AJCC 7th Edition breast cancer staging system, sensitivity and 95% Pearson-Clopper confidence intervals (CI) are reported at 90% specificity for the number of samples (N). Citation Format: Noura Tbeileh, Taylor Cavazos, Mehran Karimzadeh, Jeffrey Wang, Alice Huang, Dung Ngoc Lam, Seda Kilinc, Jieyang Wang, Xuan Zhao, Andy Pohl, Helen Li, Lisa Fish, Kimberly Chau, Marra Francis, Lee Schwartzberg, Patrick Arensdorf, Hani Goodarzi, Fereydoun Hormozdiari, Babak Alipanahi. Cell-free orphan noncoding RNAs and AI enable early detection of invasive breast cancer and ductal carcinoma in-situ [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-13-08.

Abstract PO1-08-03: Using Telehealth to Establish a High Risk Cancer Prevention Clinic over a large Geographic Area

Abstract There are health disparities in breast cancer screening based on urban, rural or frontier geographic designations. For instance, in Utah, the percentage of women who had no mammography in the last two years was 33.5% for urban, 33.8% for rural and 40.4% for frontier designations. Rural women are also diagnosed with breast cancer at later stages. Diagnosing breast cancer at earlier stages leads to improved survival, more affordable care, less toxic treatments, better quality of life and less time away from home and work. We are presenting the innovative use of telehealth to establish a High Risk Cancer Prevention Clinic to effectively reach a large geographic area and mitigate obstacles to accessing care. The clinic's mission is to identify and care for individuals with increased cancer risk due to familial, genetic and/or high risk lesions. This initiative provides centralized comprehensive evaluation, counseling and screening planning as well as referral for risk-reducing interventions closer to home. With a collaborative approach involving surgical and medical oncology, genetics, radiology and women's health specialists, the teleclinic has successfully served 280 patients since Spring 2022. Recently our health care system has increased significantly in geographic reach due to mergers with other health systems. We think this teleclinic is nimble enough and easily scalable to accommodate caring for increased numbers of patients needing this service. Citation Format: Maricel Purcell, Teresa Reading, Ginger Lee, Jennifer Tittensor, Margaret VanMeter. Using Telehealth to Establish a High Risk Cancer Prevention Clinic over a large Geographic Area [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-08-03.

Drosophila Nhe2 Over-expression Induces Autophagic Cell Death.

Autophagy is a conserved catabolic process where double membrane-bound structures form around macromolecules or organelles targeted for degradation. Autophagosomes fuse with lysosomes to facilitate degradation and macromolecule recycling for homeostasis or growth in a cell autonomous fashion. In cancer cells, autophagy is often upregulated and helps cancer cells survive nutrient deprivation and stressful growth conditions. Here, we propose that the increased intracellular pH (pHi) common to cancer cells is sufficient to induce autophagic cell death. We previously developed tools to increase pHi in the Drosophila eye via over-expression of DNhe2, resulting in aberrant patterning and reduced tissue size. We examined fly eyes at earlier stages of development and found fewer interommatidial cells. We next tested whether this decrease in cell number was due to increased cell death. We found that the DNhe2-induced cell death was caspase-independent, which is inconsistent with apoptosis. However, this cell death required autophagy genes, which supports autophagy as the mode of cell death. We also found that expression of molecular markers supports increased autophagy. Together, our findings suggest new roles for ion transport proteins in regulating conserved, critical developmental processes and provide evidence for new paradigms in growth control.

Abstract PO1-08-12: Genomic Characterization of the Carolina Breast Cancer Study

Abstract Background: Breast cancer is a heterogeneous disease defined by distinct subtypes, mutational profiles, and genomic characteristics. Previous analyses in The Cancer Genome Atlas (TCGA) have been instrumental in understanding the genomic landscape of breast cancer, including identification of key driver mutations, mutational signatures, and distinguishing features of breast cancer subtypes. However, TCGA may not be representative of other population-based breast cancer cohorts, particularly those of earlier stage and including a more diverse set of patients. Our aim was to characterize the genomic landscape of breast cancer in the Carolina Breast Cancer Study (CBCS), a cohort which oversamples Black and young women. Methods: We used targeted sequencing to profile somatic alterations in 1,175 genes from 275 formalin fixed paraffin embedded primary breast tumors in CBCS, 52% of which came from Black women, 41% from women under age 50, and 29% of which were Basal-like based on bulk RNA profiling. We also evaluated TP53 mutation as a source of breast cancer heterogeneity, comparing the type and location of TP53 mutations in each cohort. We assessed chromosome arm-level gains and losses using GISTIC and defined genomic instability as the total number of aneuploid chromosome arms. Intratumoral heterogeneity was assessed with PyClone-VI and defined as the total number and Shannon’s Diversity Index of tumor-specific subclones. For genes mutated in more than 5% of samples and chromosomal aberrations significantly enriched in GISTIC, we estimated the prevalence of alterations and compared to previous distributions in 981 TCGA tumors. Results: Seven genes [TP53 (N=101, 37%), PIK3CA (N=76, 28%), GATA3 (N=32, 12%), CDH1 (N=31, 11%), MAP3K1 (N=22, 8%), KMT2C (N=20, 7%), and CBFB (N =19, 7%)] were mutated in more than 5% of CBCS tumors; all but CBFB which were also highly prevalent in TCGA. While most TP53 hotspot mutations observed in CBCS were previously reported in TCGA, CBCS had twice as many Y220C mutations, one-third fewer R175H mutations, and a non-significantly higher proportion of nonsense mutations than TCGA (20% vs 12%, p = 0.05). Hotspot mutation prevalence in other Luminal-associated driver genes (e.g. PIK3CA, GATA3, CDH1) did not differ by dataset. CBCS tumors showed a higher degree of genomic instability (8 arms vs. 5 arms, p &amp;lt; 0.001) and subclonal diversity than TCGA (48% vs 12% comprised of two or more subclones, p &amp;lt; 0.05). Both datasets showed significant amplifications of 1q and 8q, and deletions of 13q and 17p (where BRCA2 and TP53, respectively, are located), with CBCS having non-significantly higher prevalence of each of these changes. Conclusions: While comparisons should be interpreted in light of technical and population differences between TCGA and CBCS, the overall results show that the suite of commonly altered genes and chromosome arms were highly consistent between TCGA and the diverse CBCS cohort. CBCS tumors tended to display higher genomic instability, intratumoral heterogeneity, and number and diversity of TP53 mutations. This may reflect distinctions in tumor evolutionary state between CBCS and TCGA tumors, and/or could reflect differences in the study populations. Results underscore the importance of considering population characteristics – particularly stage and race – in large-scale genomic contexts, and highlight the importance of diverse cohorts in genomic research. Citation Format: Sarah Van Alsten, Ebonee Butler, Benjamin Calhoun, Michael Love, Charles Perou, Katherine A Hoadley, Melissa Troester. Genomic Characterization of the Carolina Breast Cancer Study [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-08-12.