L1 Treatment Research Articles

Kojic Acid Derivative as an Antimitotic Agent That Selectively Kills Tumour Cells

Background/Objectives: The primary method used to pharmacologically arrest cancer development and its metastasis is to disrupt the cell division process. There are a few approaches that may be used to meet this objective, mainly through inhibiting DNA replication or mitosis. Despite intensive studies on new chemotherapeutics, the biggest problem remains the side effects associated with the inhibition of cell division in non-tumoural host cells. Methods: The efficacy and selectivity of the kojic acid derivative (L1) was studied in vitro with the use of tumoural (Caco2, SW480, HT29, T98G) and non- tumoural (HEK293T, RAW) cell lines. Light and electron microscopy observations were supported by the next generation sequencing (NGS), cytoflow, and spectroscopy analysis of mRNA and biomolecules, respectively. Results: The light and electron microscopy observations showed that L1 treatment leads to significant morphological changes in Caco2 cells, which are characteristic of mitosis arrest. Moreover, the fluorescent tubulin staining revealed the formation of tubulin ring structure associated with the apoptotic stage. Mitotic exit into apoptosis was further conformed by the cytoflow of early/late apoptosis stages and caspase-3 analysis. NGS investigation showed differentiated expressions of genes involved in mitosis and apoptosis processes. The observed IC50 in tumoural cell lines were as follows: Caco2 (IC50 = 68.2 mM), SW480 (IC50 = 15.5 mM), and HT29 (IC50 = 4.7 mM). Conclusions: The findings presented here suggest that L1 could be a valid candidate for oral prevention and/or chemotherapy in colorectal cancer. Considering high selectivity of L1 versus tumoural cell lines, more in-depth mechanistic studies could reveal unknown stages in carcinogenesis.

Investigation of the anti-Huanglongbing effects using antimicrobial lipopeptide and phytohormone complex powder prepared from Bacillus amyloliquefaciens MG-2 fermentation

Global citrus production has been severely affected by citrus Huanglongbing (HLB) disease, caused by Candidatus Liberibacter asiaticus (Clas), and the development of effective control methods are crucial. This study employed antimicrobial lipopeptide and phytohormone complex powder (L1) prepared from the fermentation broth of the endophytic plant growth promoting bacterium (PGPB) of Bacillus amyloliquefaciens strain MG-2 to treat Candidatus Liberibacter asiaticus (CLas)-infected ‘Citrus reticulata ‘Chun Jian’ plants. Real-time fluorescence quantitative polymerase chain reaction (qPCR) and PCR were employed for disease detection. The results revealed that after 15 spray-drench treatments with L1 solution, the HLB infection rate decreased from 100 to 50%, the bacterial titer decreased by 51.9% compared with a 27.9% decrease in the control group. L1 treatment triggered the production of reactive oxygen species, increased lignin content, and increased defense enzyme activities (p < 0.05). Defense-related gene expression significantly increased within 12 h of treatment. In addition, L1 application also promoted plant growth, as evidenced by higher transpiration rates and net photosynthetic rates as well as increased leave or root density. Root flora analysis revealed that the abundances of Burkholderia_thailandensis, unclassified_g_Burkholderia-Caballeronia-Paraburkholderia, unclassified_g__Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, and Pseudomonas_mosselii were 1.64, 1.46, 5.84, and 6.93 times greater, respectively, than those in the control group. The levels of phenylpropanoids, polyketides, lipids, lipid-like molecules, organic acids, and derivatives, significantly increased following L1 treatment (FC > 2, p < 0.05). Additionally, salicylic acid, dihydrojasmonic acid, and isopentenyl adenosine levels in leaves markedly increased. High-performance liquid chromatography (HPLC) confirmed that L1 contained surfactin, iturin and fengycin cyclic-lipopeptides (CLPs) as well as indole-3-acetic acid (IAA), 3-indolebutyric acid (IBA), indole-3-carboxylic acid and indole-3-carboxaldehyde auxins, N6-entopentenyladenine and t-zeatin-riboside cytokinins, abscisic acid, 1-aminocyclicpanecarboxylic acid, salicylic acid, and gibberellin A1, A3 and A4 phytohormones. These findings provide insight into multiple mechanisms by which endophytic Bacillus PGPB L1 is able to combat HLB disease, to promote citrus plant growth, and to optimize the root flora for soil health which offering an innovative strategy for sustainable management of this severe disease and improving citrus plant growth and productivity

Real-world Effectiveness of Single-Agent Enfortumab Vedotin in Patients With Advanced Urothelial Carcinoma.

Enfortumab vedotin (EV) is a nectin-4-directed antibody and microtubule inhibitor conjugate indicated for patients with metastatic urothelial carcinoma (mUC) who have received prior platinum-based chemotherapy and PD-1/L1 inhibitors or are ineligible for cisplatin-containing regimens and have undergone at least 1 prior line of therapy. EV is also indicated in combination with pembrolizumab in the first-line setting. However, real-world effectiveness of EV based on treatment line and impact of prior therapy remains unclear. This retrospective study utilized the nationwide Flatiron Health electronic health record (EHR)-derived de-identified database. Patients with advanced, recurrent, or mUC of upper or lower urinary tract who received single-agent EV in the second-line or beyond after December 18, 2019 (FDA accelerated approval date), were included. Patients without documentation of first-line therapy or without evidence of contact for 90 days from mUC diagnosis were excluded. Time to next therapy (TTNT) and overall survival (OS) were analyzed based on treatment line and prior platinum-based chemotherapy and PD-1/L1 inhibitors using Kaplan-Meier survival estimates and its 95% confidence interval. Between January 17, 2020, and September 30, 2022, 6566 patients with mUC received systemic treatment, with 431 receiving EV. EV monotherapy was administered across various lines (N = 371): second (157), third (132), fourth (62), and fifth (20). Approximately 22% of patients had prior platinum-based therapy, and 48% had prior PD-1/L1 inhibitors. Median TTNT and OS varied across treatment lines, with patients with prior platinum exposure generally showing longer TTNT and OS, notably in those receiving EV in the fourth-line setting. EV demonstrates efficacy in patients with mUC regardless of prior receipt of platinum-based chemotherapy and PD-1/L1 inhibitors or treatment line. While only hypothesis-generating, these findings provide valuable insights for patient counseling, prognostication, and therapeutic decision-making in clinical practice.

Outcome of posterior decompression by L1 laminectomy, transpedicular fixation, posterolateral fusion in burst fracture L1 with incomplete spinal cord injury

Introduction: Burst fractures are more severe than compression fractures as the bones spread out in all directions and may damage the spinal cord, leading to paralysis or nerve injury. This study aims to find the surgical outcomes of Posterior Decompression by L1 Laminectomy in terms of burst fracture with incomplete spinal cord injury. Methods: 48 adult patients with burst fractures with incomplete spinal injury came to NITR from June 2023 to May 2024. In this non-randomized clinical trial, the outcomes of decompression by L1 laminectomy were analysed by using the canal encroachment ratio, while deformity correction was assessed using the sagittal Cobb angle and the percentage of anterior vertebral height for the treatment of L1 burst fracture with incomplete spinal cord injury. Results: The patients were followed up for an average of 12 months. The mean operation time was 154.53 minutes, mean intraoperative blood loss of 48.84 mL, and the mean incision length was 7.78 cm. The canal encroachment ratio decreased significantly, as well as the sagittal Cobb angle, and anterior vertebral percentage height increased. Oswestry Disability Index, and American Spinal Injury Association impairment scale classification were significant. Conclusions: Posterior decompression combined with Transpedicular Fixation and Posterolateral Fusion in the treatment of Burst Fracture L1 with Incomplete Spinal Cord Injury is safe and effective.

7699 Unanticipated Harmony: PD-L1 Inhibitor Treatment for Urothelial Cancer Resolving Graves' Disease in a Patient - A Case of Immune Modulation Surprises

Abstract Disclosure: K. Tiwari: None. C.A. Resta: None. Introduction: Programmed death ligand 1(PD-L1) inhibitors constitute a class of immune checkpoint inhibitors (ICI) employed in treating various cancers. Thyroid-related immune-related adverse events (irAEs), such as hypothyroidism, are found to be in 3.9% of patients with PD L1 treatments. This case explores a unique scenario where a patient with pre-existing Graves' disease, undergoing PD-L1 inhibitor therapy for cancer, experienced an unexpected sequence of events leading to the default treatment of Graves' disease and subsequent progression to hypothyroidism. Case Presentation: A 59-year-old male, who had been effectively managed on methimazole for Graves' disease over two years and had chosen not to pursue definitive therapy for the condition, presented with tremors and a bilaterally enlarged thyroid gland during treatment for urothelial carcinoma. Laboratory findings included a TSH of 0.03 mIU/L (nl 0.40-4.50 mIU/L), fT4 of 3.4 ng/dL (nl 0.8-1.8 ng/dl), and tT3 of 284ng/dl (nl 76-181 ng/dl). The patient had recently started treatment with Avelumab, a PD-L1 inhibitor, as maintenance therapy following 4 cycles of gemcitabine/cisplatin cycles. Concerned about worsening Graves' disease, the methimazole dose was increased, and follow-up labs were performed after three weeks. These results showed low-normal fT4 of 0.8 ng/dL, TSH of 1.01 mIU/L(N), and low T3 of 58 ng/dL. TSI was mildly elevated at 168%. Due to rapidly changing thyroid function tests and suspicion of thyroiditis, methimazole was stopped. A PET scan a week later indicated diffuse thyroid uptake consistent with thyroiditis. Repeat labs 10 days later revealed a TSH of 67.36 mIU/L(H) and fT4 of 0.4 ng/dL(L), indicative of overt primary hypothyroidism. The patient exhibited symptoms including fatigue, constipation, and cold intolerance. The patient was initiated on levothyroxine therapy to address overt primary hypothyroidism, likely secondary to destructive thyroiditis from ICI. Discussion: Thyroid irAEs are typically incidentally discovered during routine monitoring of ICI treatment. It includes thyrotoxicosis, subclinical hypothyroidism, and overt hypothyroidism. These events commonly occur in patients lacking pre-existing thyroid conditions. In contrast, our patient, with a history of Graves' disease, experienced resolution of Graves' disease following treatment with PD-L1 inhibitor use with subsequent progression to hypothyroidism. Conclusion: This case emphasizes the importance of recognizing atypical presentations of thyroid irAEs in individuals with pre-existing thyroid disorders undergoing ICI treatment. Presentation: 6/3/2024

Autochthonous probiotic bacteria improve intestinal pathology and histomorphology, expression of immune and growth-related genes and resistance against Vibrio alginolyticus in Asian seabass (Lates calcarifer).

The study isolated two strains of intestinal autochthonous bacteria Lactiplantibacillus plantarum1 (MH155966.1) (L1) and Lactiplantibacillus plantarum2 (MH105076.1) (L2) from the Choobdeh Abadan region. The aim of this study was to investigate the effects of different strains of probiotic bacteria on the growth performance, digestive enzyme activity, histopathologic and histomorphometric characterization of the intestine, expression of immune and growth related genes, and evaluate Lates calcarifer resistance against Vibrio alginolyticus. To achieve this, for each treatment 60 L. calcarifer juveniles (75 ± 12g) were randomly distributed in three fiberglass tanks (300L) and fed for 45 days. The treatments were established as Diet 1 (control diet); L1 (diet with Lb. plantarum isolated 1); L2 (diet with Lb. plantarum isolated 2) with a bacterial concentration of 1 × 109 CFU/g. Nine fish from each treatment were sampled and examined, after euthanasia. The fish were placed 2cm from the beginning of the intestine for microscopic sampling of villi height, villi width and thickness of the epithelium, with 3 treatments: The result showed differences in the mean values of total weight were found at the end of the experiment. After 45 days of culture, the fish fed with L1 had higher (P < 0.05) growth performance than the other treatment groups. But at the end of the trial, in L2, the digestive enzyme activities were higher (P < 0.05) than the other treatment groups. The fishes fed diets supplemented with the L2 group, like the digestive enzyme activities test, presented an increase in the thickness of the epithelium of the intestine, and villus height, and villus width were greatest in L2. Fish feeding with L1 and L2 probiotics induced higher transcription levels of interleukin-10 (IL-10), granulocyte-macrophage colony-forming cells (GMCFC), epidermal growth factor (EGF), and Transforming Growth Factor Beta (TGF-β) genes in the gut, which may correlate with better immune and hematological parameters in these groups. The results of the challenge test revealed that the percentage of survival was significantly higher in L1 (76.2%) and L2 (80.95%) treatments than in the control (P < 0.05). These results indicate that host-derived probiotics (Lb. plantarum) have significant potential as important probiotics to enhance nutrient utilization, Digestive enzymes, and metabolism by increasing the gut surface area of Lates calcarifer juveniles at 45 days of culture.

Enhancing lavender (Lavandula angustifolia Mill.) propagation in vertical farming system: Investigating the impact of LED red-to-blue ratios and subsequent post-potting growth dynamics

This study aims to examine the influence of red-to-blue (R:B) LED light ratios on the propagation of Lavandula angustifolia within a vertical farming system. Additionally, it seeks to evaluate the persistent effects of LED lighting treatments on post-potting development in a glasshouse.The objective of this study is to assess how different R:B LED light ratios impact Lavandula angustifolia propagation in a vertical farming system and to analyze the enduring effects of these lighting treatments during the post-potting development stage in a glasshouse.The study consists of two phases. In the initial phase, an analysis was conducted on root emergence, root and shoot biomass, chlorophyll content, and flower bud emergence. Two LED treatments, labeled L1 and L2, were employed, with specific R:B ratios (7:1 and 9:1) and light intensities set at 140 µmol m−2 s−1 and 100 µmol m−2 s−1, respectively. Correlation and regression analyses were performed to understand the relationships among the various parameters.In Phase One, the L1 and L2 LED treatments significantly enhanced root emergence and biomass. Notably, the R:B ratios of 7:1 and 9:1, with corresponding intensities of 140 µmol m−2 s−1 and 100 µmol m−2 s−1, demonstrated substantial improvements. In the subsequent glasshouse phase, plants previously treated with L1 exhibited superior plant height and shoot biomass.The findings of this study highlight the intricate relationships among different parameters, providing valuable insights for lavender cultivation practices. The positive outcomes, particularly observed in the L1 treatment, emphasize the potential of vertical farming systems and LED lighting in optimizing lavender growth dynamics. This study underscores the importance of specific R:B ratios, contributing to sustainable propagation strategies, and acknowledges the space-efficient nature of vertical farming systems.

Cabozantinib with immune checkpoint inhibitor versus cabozantinib monotherapy in patients with metastatic clear cell renal cell carcinoma progressing after prior immune checkpoint inhibitor.

Rechallenge with antibodies targeting programmed cell death protein-1 or its ligand (PD-1/L1) after discontinuation or disease progression in solid tumors following a prior PD-1/L1 treatment is often practiced in clinic. This study aimed to investigate if adding PD-1/L1 inhibitors to cabozantinib, the most used second-line treatment in real-world patients with metastatic clear cell renal cell carcinoma (mccRCC), offers additional benefits. Using de-identified patient-level data from a large real-world US-based database, patients diagnosed with mccRCC, who received any PD-1/L1-based combination in first-line (1L) setting, followed by second-line (2L) therapy with either cabozantinib alone or in combination with PD-1/L1 inhibitors were included. Patients given a cabozantinib-containing regimen in 1L were excluded. The study end points were real-world time to next therapy (rwTTNT) and real-world overall survival (rwOS) by 2L. Of 12,285 patients with metastatic renal cell carcinoma in the data set, 348 patients met eligibility and were included in the analysis. After propensity score matching weighting, cabozantinib with PD-1/L1 inhibitors versus cabozantinib (ref.) had similar rwTTNT and rwOS in the 2L setting. Hazard ratios and 95% confidence interval (CI) for rwTTNT and rwOS are 0.74 (95% CI, 0.49-1.12) and 1.15 (95% CI, 0.73-1.79), respectively. In this study, the results align with the phase 3 CONTACT-03 trial results, which showed no additional benefit of adding PD-L1 inhibitor to cabozantinib compared to cabozantinib alone in 2L following PD-1/L1-based therapies in 1L. These results from real-world patients strengthen the evidence regarding the futility of rechallenge with PD-1/L1 inhibitors.

The Impact of Light Intensities on the Phenotypic Parameters of Cucumber Seedlings at Three Developmental Stages

Industrialized seeding production is one of the most important methods used to raise seedlings in a standardized and efficient manner. However, its consumption of light energy limits its development. In this paper, we studied the effects of LED light intensities (30 (L1), 60 (L2), 90 (L3), 120 (L4), 150 (L5), 180 (L6), and 210 (L7) µmol·m−2·s−1) on the phenotypic parameters and health index of seedlings during different growth stages (cotyledon flattening (CF), one bud and one leaf (OBOL), and one bud and two leaves (OBTL)). Ten out of sixteen plants were selected to measure the parameters in every treatment. The control group was treated with 150 µmol·m−2·s−1 all the time. The results show that the height of the L1-treated plants was greatest during the cotyledon flattening stage. The L7 treatment produced the largest stem diameter, the greatest biomass, the largest leaf area, and the largest leaf circumference, the values of which were 2.61 mm, 3.3 g, 60.67 cm2, and 39.71 cm, respectively. During the one bud and one leaf stage, the L1 treatment produced the tallest plants. The L7 treatment resulted in the largest stem diameter, the greatest biomass, the largest leaf area, and the largest leaf circumference. During the one bud and two leaves stage, the plant height, stem diameter, leaf circumference, and biomass produced of the L6 treatment were the greatest, with values of 68.37 mm, 3.09 mm, 51.54 cm, and 4.47 g. L5 resulted in the largest leaf area (63.91 cm2). The seedling index of the three stages showed an inflection point at the L4 level. The experimental results showed that the light intensity required by the cucumber seedlings was 120 µmol·m−2·s−1, 150 µmol·m−2·s−1, and 120 µmol·m−2·s−1 during the three stages, from the cotyledon flattening stage to the one bud and two leaves stage. The results provide references regarding the light required during the cultivation of cucumber seedlings.

Effect of Light Quality on Seed Potato (Solanum tuberose L.) Tuberization When Aeroponically Grown in a Controlled Greenhouse.

A plant factory equipped with artificial lights is a comparatively new concept when growing seed potatoes (Solanum tuberosum L.) for minituber production. The shortage of disease-free potato seed tubers is a key challenge to producing quality potatoes. Quality seed tuber production all year round in a controlled environment under an artificial light condition was the main purpose of this study. The present study was conducted in a plant factory to investigate the effects of distinct spectrum compositions of LEDs on potato tuberization when grown in an aeroponic system. The study was equipped with eight LED light combinations: L1 = red: blue: green (70 + 25 + 5), L2 = red: blue: green (70 + 20 + 10), L3 = red: blue: green (70 + 15 + 15), L4 = red: blue: green (70 + 10 + 20), L5 = red: blue: far-red (70 + 25 + 5), L6 = red: blue: far-red (70 + 20 + 10), L7 = red: blue: far-red (70 + 15 + 15), L8 = red: blue: far-red (70 + 10 + 20), and L9 = natural light with 300 µmol m-2 s-1 of irradiance, 16/8 h day/night, 65% relative humidity, while natural light was used as the control treatment. According to the findings, treatment L4 recorded a higher tuber number (31/plant), tuber size (>3 g); (9.26 ± 3.01), and GA3 content, along with better plant growth characteristics. Moreover, treatment L4 recorded a significantly increased trend in the stem diameter (11.08 ± 0.25), leaf number (25.32 ± 1.2), leaf width (19 ± 0.81), root length (49 ± 2.1), and stolon length (49.62 ± 2.05) compared to the control (L9). However, the L9 treatment showed the best performance in plant fresh weight (67.16 ± 4.06 g) and plant dry weight (4.46 ± 0.08 g). In addition, photosynthetic pigments (Chl a) (0.096 ± 0.00 mg g-1, 0.093 ± 0.00 mg g-1) were found to be the highest in the L1 and L2 treatments, respectively. However, Chl b and TCL recorded the best results in treatment L4. Finally, with consideration of the plant growth and tuber yield performance, treatment L4 was found to have the best spectral composition to grow quality seed potato tubers.

The Effects of Shading on the Photosynthetic Performance of Endangered Plant Horsfieldia hainanensis Seedlings

Shading is one of the management practices for preventing the damage or injury of plant seedlings during extreme weather and climate events, such as very high temperatures and heat stress. In this study, we investigated the effects of different shading conditions on the photosynthetic characteristics of the endangered plant Horsfieldia hainanensis in Guangxi, China. The H. hainanensis seedlings in this study underwent five shading treatments, including 20% (L1), 40% (L2), 60% (L3), 80% (L4), and 100% (control) of full sunlight. The net growth of their diameter and height, and photosynthetic gas exchange parameters including their photosynthesis rate (Pn), transpiration rate (Tr), intercellular CO2 concentration (Ci), and water use efficiency (WUE) were measured for the examined seedlings. The OJIP curve and 820 nm light absorption curve, and the osmotic substances and products of membrane lipid peroxidation were employed to assess photosynthetic capacity, identify the factors constraining photosynthetic carbon assimilation, and investigate the mechanisms influencing photosystem II (PSII) and photosystem I (PSI) in the seedlings under shade stress. The results showed that the seedlings in the L2 treatments had the highest net growth and Pn, the best photosynthetic performance, and the best coordination between PSII and PSI. The net photosynthesis (Pn) levels exhibited a declining trend in the following order: L2 &gt; L3 &gt; L4 &gt; L1. In the L1 treatment, non-stomatal factors emerged as the primary determinant affecting the Pn of the seedlings. The performance index (potential) of PSII, representing the conservation of absorbed photon energy to intersystem electron acceptor reduction (PIABS and ΔI/I0) of the seedlings, decreased in the order of L2 &gt; L3 &gt; L4 &gt; L1. The photosystem performance and the coordination between PSII and PSI (Φ(PSI/PSII)) of the seedlings decreased in the order of L2 &gt; L1 &gt; L3 &gt; L4. Under the low and moderate shading stresses (L1–L3), more serious damages occurred in PSII than in PSI, including on the donor side of PSII and in the electron transfer from QB to the acceptor side of PSI. In contrast, more considerable injury occurred in PSI than in PSII under the stress of the heavy shading treatment (L4). Considering the alterations in their leaf osmotic regulatory substances and membrane lipid peroxidation products, our findings indicate that the L2 treatment was the most conducive to the growth of the H. hainanensis seedlings. In contrast, the L1 treatment subjected H. hainanensis seedlings to the most significant stress, resulting in substantial damage to their growth and photosynthetic mechanisms. Our research provides a scientific insight into and a practical guide for the selection of an appropriate light intensity for the conservation and cultivation of endangered plant species, such as H. hainanensis.

Temporal changes in treatment patterns by age group and functional status before and after PD-1/L1 inhibitor approvals in advanced urothelial carcinoma.

Metastatic urothelial carcinoma (mUC) has poor prognosis. A high unmet need exists for novel treatment for those who are unfit for platinum-based chemotherapy. We aimed to describe real-world temporal changes in patient characteristics and 1L treatment selection for mUC patients in the United States following the approval of anti-PD-1/L1 treatments. This study was a retrospective, observational study using anonymized and structured oncology electronic medical record (EMR) data from IQVIA and the US Oncology Network iKnowMed (USON). After approval of 1L anti-PD-1/L1 treatment for mUC, there is a marked increase in the use of 1L anti-PD-1/L1 monotherapies, accompanied by a proportional decrease in 1L platinum-based treatments and non-guideline-based therapy; particularly among the elderly (> 75 years) and those with poor ECOG performance status (ECOG PS 2+). Anti-PD-1/L1 monotherapies fulfill the prior unmet need of frail mUC patients who are ineligible for platinum-based therapies.

Effect of isolated lactic acid bacteria on the quality and bacterial diversity of native grass silage.

The objective of this study was to isolate lactic acid bacteria (LAB) from native grasses and naturally fermented silages, determine their identity, and assess their effects on silage quality and bacterial communities of the native grasses of three steppe types fermented for 60 days. Among the 58 isolated LAB strains, Limosilactobacillus fermentum (BL1) and Latilactobacillus graminis (BL5) were identified using 16S rRNA sequences. Both strains showed normal growth at 15- 45°C temperature, 3-6.5% NaCl concentration, and pH 4-9. Two isolated LAB strains (labeled L1 and L5) and two commercial additives (Lactiplantibacillus plantarum and Lentilactobacillus buchneri; designated as LP and LB, respectively) were added individually to native grasses of three steppe types (meadow steppe, MS; typical steppe, TS; desert steppe, DS), and measured after 60 d of fermentation. The fresh material (FM) of different steppe types was treated with LAB (1 × 105 colony forming units/g fresh weight) or distilled water (control treatment [CK]). Compared with CK, the LAB treatment showed favorable effects on all three steppe types, i.e., reduced pH and increased water-soluble carbohydrate content, by modulating the microbiota. The lowest pH was found in the L5 treatment of three steppe types, at the same time, the markedly (p < 0.05) elevated acetic acid (AA) concentration was detected in the L1 and LB treatment. The composition of bacterial community in native grass silage shifted from Pantoea agglomerans and Rosenbergiella nectarea to Lentilactobacillus buchneri at the species level. The abundance of Lentilactobacillus buchneri and Lactiplantibacillus plantarum increased significantly in L1, L5, LP, and LB treatments, respectively, compared with CK (p < 0.05). In summary, the addition of LAB led to the shifted of microbiota and modified the quality of silage, and L. fermentum and L. graminis improved the performance of native grass silage.

Phase I/II combination study of tifcemalimab with toripalimab in patients with refractory extensive stage small cell lung cancer (ES-SCLC).

8579 Background: The B- and T-lymphocyte attenuator (BTLA) is a novel inhibitory co-signaling receptor expressed on B cell, T cells and NK cells. Co-blockade of the BTLA and PD-1 pathways improved antigen specific anti-tumor T cell response. Tifcemalimab (JS004 or TAB004) is a humanized IgG4 monoclonal antibody with a hinge mutation (S228P) that binds BTLA and blocks its interaction with its ligand HVEM. In previous phase I studies, tifcemalimab has shown preliminary anti-tumor activities as monotherapy or in combination with toripalimab (anti-PD-1) with a manageable safety profile in patients with advanced malignancies. Methods: Eligible ES-SCLC patients refractory to prior therapies were enrolled in this I/II study (NCT05000684). Patients received 200mg tifcemalimab and 240mg toripalimab intravenously once every three weeks until disease progression, intolerable toxicity or 2 years of treatment. Study objectives included safety, anti-tumor activity and correlative biomarkers. Results: As of Jan 31, 2023, a total of 43 ES-SCLC patients refractory to prior therapy were enrolled. The median age was 60.0 (range 38-75) years. The median prior line of therapy was 1 and 14 (32.6%) patients received prior anti-PD-1/L1 treatment. By the cut-off date, the median follow-up duration was 12.1 weeks. Thirty-two (74.4%) patients experienced treatment-emergent adverse events (TEAEs); 12 (27.9%) patients experienced grade ≥ 3 TEAEs. The most common TEAEs were hyponatraemia (16.3%), alanine aminotransferase increased (14%), aspartate aminotransferase increased (14%), and blood creatine phosphokinase increased (14%). Three (7.0%) patients experienced treatment-related adverse events (TRAEs) led to interruption of study drugs and no TRAEs led to discontinuation of study drugs were reported. Fifteen (34.9%) patients experienced immune related AE (irAEs), and 2 (4.7%) patients experienced grade ≥ 3 irAEs. Among 38 efficacy evaluable patients, the ORR was 26.3% and the DCR was 57.9%. The ORR was 8.3% in immunotherapy treated patients and 40.0% in immunotherapy naïve patients. By the cut-off date, 70.0% of the responses were ongoing and the median duration of response was not reached. Tumor expression of HVEM and PD-L1 was evaluated to explore the correlation with clinical response. Conclusions: Tifcemalimab in combination with toripalimab were well tolerated in patients with refractory ES- SCLC. Further clinical evaluation of this combination treatment in SCLC is warranted. Clinical trial information: NCT05000684 .

SI-B003 (PD-1/CTLA-4) in patients with advanced solid tumors: A phase I study.

e14668 Background: Although marketed antibodies against PD-1/L1 or CTLA-4 have been shown to be efficacious in a variety of tumors, the overall response rate (ORR) remains low. SI-B003 is a PD-1/CTLA-4 bispecific antibody. We hypothesized that co-targeting PD-1 and CTLA-4 simultaneously could achieve a better anti-tumor activity due to cooperative binding to exhausted T-cell subsets. Methods: Patients with recurrent or metastatic solid tumors who had failed standard therapy were eligible to participate. SI-B003 was administered intravenously every other week (Q2W) in a four-week cycle. The phase Ia dose escalation consisted of an accelerated titration phase (the first 2 dose levels) and a mTPI-2 with a total of five dose levels: 0.1, 0.3→1.0, 3, 5 and 10 mg/kg. The phase Ib consisted of a dose expansion and indication expansion cohorts with 5, 10 mg/kg Q2W and 7.5mg/kg Q3W. The primary objectives of phase Ia were to assess dose-limiting toxicity (DLT), maximum tolerated dose (MTD) or maximum administration dose (MAD), and treatment emergent adverse events (TEAEs). The primary objectives of phase Ib included safety, tolerability, and identification of the recommended phase 2 dose (RP2D). Results: As of December 31, 2022, 60 patients with solid tumors received SI-B003 at six different dose levels (14 in Ia, 46 in Ib). Median age was 55.5 years. The median PFS (95%CI) was 3.7 (1.6~5.7) months. Among 56 evaluable patients, the objective response rate (ORR) (n/N, 95%CI) was 16.1% (9/56, 7.6~28.3%) and disease control rate (DCR) was 50.0% (28/56). Confirmed ORR, DCR, median duration of response (DoR) and median progression free survival (PFS) were 9.5% (2/21, 1.2~30.4%), 42.9% (9/21), NR and 2.1 (95%CI, 1.4~NA) months in the 5mg/kg Q2W group and 8.3% (2/24, 1.0~27.0%), 62.5% (15/24), 7.1 (95%CI, 1.9~NA) and 5.7 (95%CI, 1.9~8.7) months in 10mg/kg Q2W group respectively. One patient received 7.5mg/kg Q3W had a confirmed response and was still under treatment. Among 21 evaluable patients who had received prior PD-1/L1 treatment, the ORR and DCR were 14.3% (3/21, 3.1~36.3%) and 52.4% (11/21). The mDoR (95%CI) and mPFS (95%CI) were 4.2 (NA) months and 4.1 (1.4, 5.7) months. The most common TRAEs were rash (35%), AST increased (28%), ALT increased (28%). The most common grade ≥3 TRAEs were AST increased (3%). One patient died from an immune-mediated pneumonia. Conclusions: SI-B003 demonstrated a tolerable safety profile and preliminary anti-tumor activity in patients with recurrent or metastatic solid tumors. Anti-tumor effects of SI-B003 were observed in patients who had progressed on prior PD-1/L1 immunotherapy suggesting that SI-B003 may be beneficial for patients who have developed resistance to PD-1/L1 drugs. Clinical trial information: NCT04606472 .

IMM01 plus tislelizumab in patients with advanced solid tumors and lymphoma: An open-label, multicenter, phase 1b/2 dose escalation and expansion study (IMM01-04).

2600 Background: IMM01, a recombinant SIRPα-Fc fusion protein, can activate macrophages to enhance anti-tumor activity by blocking CD47-SIRPα interaction. Based on preclinical results, IMM01 showed unique property of weak human erythrocyte conjugation. IMM01 plus tislelizumab has the potential to augment both innate and adaptive anti-tumor immune responses. Methods: This study included the phase 1b dose escalation part and the phase 2 dose expansion part. The dose escalation part was conducted with a standard 3+3 design to evaluate the safety, MTD/RP2D and anti-tumor activity in patients with advanced solid tumors. IMM01 (1.0, 1.5, 2.0 mg/kg) was administered once a week and tislelizumab (200mg) was administered once every 3 weeks. The results of the dose escalation part were reported here. Results: As of 26 Dec 2022, 14 pts were enrolled in the dose escalation part, including 5 NSCLC, 2 SCLC, 2 ESCC, 2 CRC, 1 HCC, 1 melanoma and 1 cervical carcinoma. Median age was 61 years (range 49-70). Median of prior therapies was 4 (range 1-10), and 13 pts (92.9%) received previous anti PD-1/L1 treatment. Treatment-related adverse events (TRAEs) occurred in 14 pts (100.0%) to date. Most TRAEs were grade 1-2. The most common TRAEs (≥10%) of all grades were anaemia (85.7%), platelet count decreased (28.6%), asthenia (28.6%), white blood cell count decreased (21.4%), lymphocyte count decreased (21.4%), IRR (21.4%), proteinuria (21.4%), gamma-glutamyltransferase increased (14.3%), white blood cell count increased (14.3%), blood creatine phosphokinase MB increased (14.3%), neutrophil count decreased (14.3%), neutrophil count increased (14.3%), hypoalbuminaemia (14.3%) and hyponatraemia (14.3%). The Grade 3-4 TRAEs were lymphocyte count decreased (21.4%), white blood cell count decreased (14.3%), platelet count decreased (14.3%), blood creatine phosphokinase MB increased (7.1%), blood pressure increased (7.1%) and anaemia (7.1%). No DLT occurred. No TRAE leading to treatment discontinuation was reported. 2 pts (14.3%) had treatment related SAE (2 platelet count decreased), they recovered without platelet transfusion and no bleeding occurred. In 14 pts, 1 pt had confirmed PR and 3 pts had SD. One heavily pre-treated NSCLC pt in the 2.0 mg/kg dose cohort had confirmed PR at Week 12, who was resistant to anti PD-1. One NSCLC pt in 1.5 mg/kg dose cohort and 2 pts (1 SCLC, 1 CRC) in 2.0 mg/kg dose cohort had SD. The treatments of these four pts are ongoing. Conclusions: Up to 2mg/kg IMM01 plus tislelizumab is well tolerated. The RP2D of IMM01 was determined as 2mg/kg, this was the highest IMM01 dose evaluated in human as a single agent. IMM01 plus tislelizumab showed preliminarily anti-tumor activity in patients with advanced solid tumors. The dose expansion part in advanced solid tumors and lymphoma is ongoing. Clinical trial information: chictr20220791 .

A phase 2 open-label study of conditionally active biologic ozuriftamab vedotin (BA3021) in failed PD-1/L1 treatment of patients with recurrent or metastatic squamous cell carcinoma of the head and neck.

TPS6107 Background: Ozuriftamab vedotin (BA3021) is a conditionally active biologic (CAB) anti-receptor tyrosine kinase orphan receptor 2 (ROR2) humanized monoclonal antibody (IgG1) conjugated to monomethyl auristatin E (MMAE) using a cleavable linker (CAB ROR2 ADC) (Short, et al., 2014, Chang, et al. 2021). Conditional and reversible binding by CABs is designed to reduce off-tumor toxicity and immunogenicity, avoid tissue-mediated drug deposition, and improve pharmacokinetics (PK). ROR2 is a cell-surface transmembrane receptor protein tyrosine kinase highly expressed in several tumor types including head and neck squamous cell cancer (HNSCC). Increased ROR2 expression has been associated with tumor resistance to chemotherapy, programmed death-1 (PD-1) inhibitors, molecular targeted therapy, and radiation therapy. A pronounced unmet need exists as the majority of patients with recurrent or metastatic HNSCC experience disease progression with existing therapies (Chow 2020). Methods: BA3021-002 is a multi-center, open-label, single-arm Phase 2 study (n=40) designed to evaluate the efficacy and safety of BA3021 in patients previously treated with PD-(L)1 with ROR-2 expressing recurrent or metastatic HNSCC. Eligible patients include those with measurable disease by Response Evaluation Criteria in Solid Tumors (RECIST) Version 1.1 criteria. Failure to prior treatment is defined as documented disease progression after no more than one PD-1/L1 inhibitor either administered as monotherapy or in combination therapy. Pharmacokinetic (PK), pharmacodynamic (PD), immunogenicity, and biomarker assessments will be performed. Enrollment is ongoing (NCT05271604)

ORGANOLEPTIC TESTS OF PEMPEK WITH VARIOUS SURIMI FORMULATIONS OF SANGKURIANG CATFISH (Clarias gariepsinus)

This study aims to study various surimi formulations of Sangkuriang catfish (Clarias gariepsinus) for the resulting pempek, using the experimental method using a Randomized Block Design (RBD) arranged in a non-factorial manner with one treatment of various formulations of surimi Sangkuriang catfish (Clarias gariepsinus) consisting of of six factors and repeated four times. Pempek organoleptic test results with hedonic tests on color, aroma, and taste. The highest preference value for the color of pempek made from sangkuriang catfish surimi in the L1 treatment (0.25 parts of sangkuriang catfish surimi and 1.00 parts of tapioca flour) with a clean white pempek color typical of fish pempek with an average value of 4.30 (criterion panelists liked). The highest preference value for the taste and aroma of pempek made from sangkuriang catfish surimi was found in treatment L6 (1.50 parts of sangkuriang catfish surimi and 1.00 parts of tapioca flour) with a savory taste and dominant pempek aroma with an average value of 3 .90 (panelists preferred criteria) and 4.30 (panelists preferred criteria).

Abstract 674: CRISPR and drug screens identify ERK as the mediator of IFNg-induced melanoma growth inhibition

Abstract Interferon-gamma (IFNg) is critical for the success of immune checkpoint blockade therapy. Disruption of IFNg signaling makes tumors resistant to anti-PD-1/L1 treatment primarily by evasion of the growth inhibition (GI) induced by this cytokine. However, the pathways mediating IFNg-induced GI are largely unknown. We used two independent approaches to delineate the IFNg GI pathway, a chemical genomics screen and a whole genome targeting CRISPR/Cas9 screen in an IFNg GI-sensitive patient-derived melanoma (PDM) cell line. Drug screening revealed that treatment with RAF and ERK inhibitors disrupted IFNg GI in PDM cells. For the CRISPR screen, gene-level analysis showed that the magnitude of enrichment for guide RNAs targeting ERK2 in IFNg-treated cells was comparable to proximal IFNg signaling proteins like JAK1, STAT1, IFNGR2, and JAK2 (&amp;gt;26-fold enrichment for ERK2, p = 2.28e-07). Thus, results from both these experiments converged upon ERK as the primary downstream target of IFNg essential for GI induction. We validated the involvement of ERK by detecting a sustained increase in phospho-ERK1/2 levels following IFNg treatment. In live imaging experiments, we found that blocking ERK activity with the ERK inhibitor Ulixertinib blocks the induction of cell death after IFNg treatment in 17 of 23 (~74%) IFNg-sensitive PDM lines covering all the MAPK mutant and triple wildtype molecular subtypes of melanoma. Decreasing and increasing ERK activity levels using shRNA and overexpression approaches also led to a decrease and increase in IFNg-mediated GI, respectively. We have identified NOXA and DR5 as likely mediators of cell death through the IFNg-ERK GI pathway by comparing gene expression profiles of control and IFNg-treated PDM cells. Finally, phorbol 12-myristate 13-acetate (PMA), a known ERK activator, synergizes with IFNg treatment to induce increased GI in PDM cells (Bliss synergy score 16.87, p = 1.83e-123). In summary, the IFNg-ERK signaling axis mediates cell death downstream of IFNg treatment in melanoma cells. This pathway is active in all melanoma subtypes, making it an attractive target to enhance IFNg GI in tumor cells. Our results provide a new understanding of the IFNg GI pathway that will also be crucial to define mechanisms of GI resistance in tumor cells. Citation Format: Ameya S. Champhekar, Rachel Heymans, Cynthia Gonzalez, Guillem Turon Font, Daniel Karin, June Lee, Robert Damoiseaux, Antoni Ribas. CRISPR and drug screens identify ERK as the mediator of IFNg-induced melanoma growth inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 674.

Association between progression-free survival and overall survival in women receiving first-line treatment for metastatic breast cancer: evidence from the ESME real-world database

BackgroundOverall survival (OS) is the gold standard endpoint to assess treatment efficacy in cancer clinical trials. In metastatic breast cancer (mBC), progression-free survival (PFS) is commonly used as an intermediate endpoint. Evidence remains scarce regarding the degree of association between PFS and OS. Our study aimed to describe the individual-level association between real-world PFS (rwPFS) and OS according to first-line treatment in female patients with mBC managed in real-world setting for each BC subtype (defined by status for both hormone-receptor [HR] expression and HER2 protein expression/gene amplification).MethodsWe extracted data from the ESME mBC database (NCT03275311) which gathers deidentified data from consecutive patients managed in 18 French Comprehensive Cancer Centers. Adult women diagnosed with mBC between 2008 and 2017 were included. Endpoints (PFS, OS) were described using the Kaplan–Meier method. Individual-level associations between rwPFS and OS were estimated using the Spearman’s correlation coefficient. Analyses were conducted by tumor subtype.Results20,033 women were eligible. Median age was 60.0 years. Median follow-up duration was 62.3 months. Median rwPFS ranged from 6.0 months (95% CI 5.8–6.2) for HR-/HER2 − subtype to 13.3 months (36% CI 12.7–14.3) for HR + /HER2 + subtype. Correlation coefficients were highly variable across subtypes and first-line (L1) treatments. Among patients with HR − /HER2 − mBC, correlation coefficients ranged from 0.73 to 0.81, suggesting a strong rwPFS/OS association. For HR + /HER2 + mBC patients, the individual-level associations were weak to strong with coefficients ranging from 0.33 to 0.43 for monotherapy and from 0.67 to 0.78 for combined therapies.ConclusionsOur study provides comprehensive information on individual-level association between rwPFS and OS for L1 treatments in mBC women managed in real-life practice. Our results could be used as a basis for future research dedicated to surrogate endpoint candidates.