bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5495: PHI-501, a potent and novel inhibitor of NRAS mutated acute myeloid leukemia
Authors: Nam Ky-Youb , Park Jung Hee , Kim Kyung Ah , Shin Inje , Han June H-J , Kim Kyu-Tae , Yoon Jeong Hyeok , Cho Hanna , Choi Seung-Hye , Sengupta Sandip , Sim Taebo
Published: June 2022

Purpose: Development of the first-in-class inhibitor of NRAS mutated acute myeloid leukemia. PHI-501 has potent activity against receptor tyrosine kinases and intracellular kinases related to STAT, AKT, and ERK signaling.

Description: Preclinical testing of PHI-501 for efficacy against NRAS mutations both in vitro and in vivo xenografts.

Summary: Mutations in the RAS genes (including KRAS, NRAS, and HRAS) are discovered in about 30% of all tumors. KRAS is the most frequently mutated gene in cancers found in pancreatic, colon, and lung, while NRAS mutations around the hot spots at codons 12, 13, and 61 are more common in AML (10.3%). PHI-501 is a novel small molecule inhibitor intended for the treatment of AML in patients expressing the N-RAS activating mutation. PHI-501 presented higher selectivity within a panel of 355 protein kinases. PHI-501 exerts their anti-proliferative effects by inducing apoptosis and G0-G1 arrest and significantly induce apoptotic markers (cleaved PARP and caspase 3) in both Ba/F3-NRAS-G12D and OCI-AML3 cells. The results of Western blot analysis showed that PHI-501 attenuate phosphorylation of p70S6K1, AKT, and p38 in OCI-AML3 After 21 days of the treatment in subcutaneous OCI-AML3 human AML xenograft model, PHI-501 showed significant inhibition of tumor growth (TGI) in a dose-dependent manner. Compared to the vehicle group, the 40 mg/kg group demonstrated a 74.0% lower average tumor volume (P<0.0001) and a 78.5% lower average tumor weight (P=0.0041). No significant body weight changes were observed in any of the groups. In toxicology and pharmacokinetic study, PHI-501 exhibited pharmacokinetic profiles favorable for oral administration in rodent, dog, and monkey and displayed acceptable in vivo and in vitro safety profiles. These results show the significant ability of PHI-501 to inhibit the progression of NRAS activated AML tumor growth, making it a strong candidate for the treatment of AML.

Conclusion: The preclinical evaluation of PHI-501, a novel N-RAS inhibitor, showed clear evidence of anticancer activity for AML and improved efficacy in both in vitro and in vivo models. Consequently, PHI-501 is a new potent multi-kinase inhibitor with characteristics that warrant entry into human trials for the treatment of AML in patients expressing the N-RAS activating mutation.

Citation Format: Ky-Youb Nam, Jung Hee Park, Kyung Ah Kim, Inje Shin, June H-J Han, Kyu-Tae Kim, Jeong Hyeok Yoon, Hanna Cho, Seung-Hye Choi, Sandip Sengupta, Taebo Sim. PHI-501, a potent and novel inhibitor of NRAS mutated acute myeloid leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5495.
Abstract 5506: CD30 co-stimulatory domain enhances the efficacy of chimeric antigen receptor-engineered γδT cells
Authors: Cho Hyun-Il , Kang Chung-Hyo , Lee Sang-Eun , Song In-Sil , Ha Jung-Min , Lee Seon-Duk , Sohn Hyun-Jung , Kim Tai-Gyu
Published: June 2022

Adoptive transfer of immune killer cells genetically engineered to express chimeric antigen receptors (CARs) has demonstrated a feasible and effective approach against numerous types of cancers. Among the killer cells, γδT cells possessing a wide range of antigen-independent tumor recognition and low risk of developing GVHD offer a chance to develop CAR-engineered allogenic ‘off-the-shelf’ cellular therapeutics. Successful tumor eradication primarily depends on providing optimized signals affecting the proliferation, persistence, antitumor activity and cytokine secretion of CAR-γδT cells. We describe here a novel CD30 costimulatory domain for CAR signaling, which is known to be involved in cell anti-apoptosis, activation and proliferation. Conventional αβT cells and innate γδT cells activated by a CD30-ζ signaling-CAR displayed similar levels of cytolytic efficacies and cytokine secretion against antigen-expressing cancer cell lines as those of other CAR containing commonly used costimulatory domains CD28, CD27, 4-1BB, OX40, and ICOS. Furthermore, the third-generation CARs including the CD30 costimulatory domain retained potent antitumor efficacies independent of their position or type of partner costimulatory domain. More significantly, CAR-γδT cells containing CD30-ζ signaling revealed effective cytolytic activities against several solid cancer cell lines based on its antigen-recognizing antibody fragments. Our findings suggest that the CD30 costimulatory domain could be an attractive candidate for developing CAR-engineered killer cell therapeutics and may be applicable for the design of numerous antigen-recognizing CAR constructs, with an emphasis on targeting solid tumors.

Citation Format: Hyun-Il Cho, Chung-Hyo Kang, Sang-Eun Lee, In-Sil Song, Jung-Min Ha, Seon-Duk Lee, Hyun-Jung Sohn, Tai-Gyu Kim. CD30 co-stimulatory domain enhances the efficacy of chimeric antigen receptor-engineered γδT cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5506.
Abstract 5520: AK129, an anti-PD1/LAG-3 bi-specific antibody for cancer therapy
Authors: Huang Zhaoliang , Pang Xinghua , Zhong Tingting , Jin Chunshan , Chen Na , Xia Dennis , Zhang Peng , Wang Max , Xia Michelle , Li Baiyong
Published: June 2022

Background: AK129 is a bi-specific antibody acts by co-blocking two immune checkpoint molecules co-expressed on T cells, PD-1 and LAG-3. Signaling through PD-1/PD-L1 exerts major effects on cytokines production by T cells, inhibiting production of IFN-γ and IL-2. LAG-3 is a cell surface molecule expressed on effector T cells and Tregs, which plays a vital role in regulating T cell function to promote tumoral immune escape. LAG-3 and PD-1 mediate different signaling pathways, but they may act synergistically to cause effector T cell inactivation and exhaustion. AK129 is being investigated as a cancer immunotherapeutic agent.

Methods: The antigen binding activity of AK129 to PD-1 and LAG-3 were measured by ELISA and FACS. In reporter gene assay, the bioactivities of AK129 to block the interaction of PD-1 with PD-L1 and to neutralize LAG-3-mediated activity were determined. The bioactivity of AK129 to enhance IL-2 and IFN-γ production in PBMCs via blockade of PD-1 and PD-L1/2 interaction were determined by using cellular assays. In in-vivo pharmacology studies, anti-tumor activity of AK129 was investigated in BALB/c-hPD1/hLAG3 mice bearing CT26.WT tumor model.

Results: AK129 showed good antigen binding to PD-1 and LAG-3. AK129 potently inhibited the binding of LAG-3 to its ligand MHC-II, and the binding of PD-1 to its ligand PD-L1, thus blocking downstream immune suppression. At the individual target level, AK129 demonstrated a similar PD-1 blocking activity compared to penpulimab, and similar LAG-3 blocking activity compared to relatlimab. Importantly, at the whole cell level, the enhancement effect of AK129 is stronger than the combination of penpulimab and relatlimab, as evidenced by significantly higher level of IL-2 and IFN-γ secretion from PBMCs. AK129 showed favorable anti-tumor activity in BALB/c-hPD1/hLAG3 mice bearing CT26.WT tumor model, with tumor growth inhibition values reaching 99.27% and 89.48% in high dose (20 mg/kg) and low dose (4 mg/kg) group of AK129, respectively.

Conclusion: AK129 is a dual checkpoint blocking bispecific antibody with design features similar to the PD-1/CTLA-4 bispecific antibody AK104, which demonstrated efficacy similar to PD-1 and CTLA-4 antibody combo and much improved safety profile. AK129 has demonstrated robust anti-tumor activity in both in vivo and in vitro pre-clinical studies, and with enhanced T cell activity compared to PD-1 and LAG-3 antibody combo. The efficacy and safety of AK129 await testing in the clinic.

Citation Format: Zhaoliang Huang, Xinghua Pang, Tingting Zhong, Chunshan Jin, Na Chen, Dennis Xia, Peng Zhang, Max Wang, Michelle Xia, Baiyong Li. AK129, an anti-PD1/LAG-3 bi-specific antibody for cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5520.
Abstract 5529: In vivo efficacy evaluation of ASC61, an oral PD-L1 inhibitor, in two tumor mouse models
Authors: Wu Jinzi J. , He Handan
Published: June 2022

Background: ASC61 is an oral inhibitor prodrug of programmed cell death-ligand 1 (PD-L1). ASC61 is converted to its pharmacologically active metabolite ASC61-A in the plasma. In vitro studies showed that ASC61-A could activate PD-L1 dimerization and membrane PD-L1 protein internalization, interfere PD-1/PD-L1 interaction and enhance T-cell activation. Here we report the in vivo efficacy of ASC61 in two tumor mouse models. ASC61 was found to have the best antitumor activity among all test drugs in the syngeneic tumor mouse model, BALB/c mice bearing CT-26-hPD-L1 tumors. Then a humanized mouse model, PD-1/PD-L1 dKI HuGEMM bearing hPD-L1 MC38 tumor, was used to further validate the therapeutic efficacy of ASC61.

Methods: In the BALB/c mouse model, mice were inoculated subcutaneously with CT-26-hPD-L1 cells. When the average tumor volume reached approximately 69 mm3, mice were randomly assigned and treated twice a day (BID) with vehicle, reference drug (Atezolizumab), ASC61, GLC01-589 or GLC01-633 for 19 days. In the PD-1/PD-L1 dKI HuGEMM mouse model, MC38 hPD-L1 cells were subcutaneously inoculated in the right rear flank region of mice. Once the mean tumor size reached approximately 78.3 mm3, mice were randomized and treated BID for 16 days with vehicle, Atezolizumab, ASC61 (50 or 100 mg/kg). In both models, body weights and tumor volumes were measured regularly, and tumor growth inhibitions (TGI) of different groups were compared.

Results: All mice showed a gradually increase in body weight during the study and no significant difference of body weight or body weight change was found among different groups in both mouse models. In the BALB/c mouse model, ASC61 50 mg/kg showed significantly inhibitory effects on the tumor growth with the best TGI value of 52.9% (p < 0.05) on Day 19, even better than that of the reference drug (40.77%). In the PD-1/PD-L1 dKI HuGEMM mouse model, Group 2 to 4 showed TGI values of 69.62%, 33.62% and 63.15% on Day 16, respectively. Atezolizumab 5 mg/kg BIW and ASC61 100 mg/kg BID produced similar significant antitumor efficacies (both p < 0.001). Survival analysis showed significant differences between Group 1 and Group 2 to 4 (p = 0.0002, 0.0338 and 0.0027, respectively).

Conclusion: Results of this study show that ASC61 is well tolerated in both mouse models, and ASC61 has comparable antitumor efficacies as the FDA-approved PD-L1 antibody, Atezolizumab. ASC61 is an oral PD-L1 inhibitor, and thus will have better patient compliance than injective PD-1/PD-L1 antibody. ASC61 first in human clinical trial will begin soon.

Citation Format: Jinzi J. Wu, Handan He. In vivo efficacy evaluation of ASC61, an oral PD-L1 inhibitor, in two tumor mouse models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5529.
Abstract 5532: An ‘anti-PD1-IL2 beta-only super-agonist’ displays potent anti-tumor efficacy
Authors: To Minh D. , Munegowda Manjunatha Ankathatti , Merchant Rosemina , Sharma Aanchal
Published: June 2022

Introduction: Combination studies with engineered IL-2 variants and immune-checkpoint blockade have demonstrated synergy in both pre-clinical models and the clinic. Co-administration of those 2 agents can potentially face challenges related to overlapping toxicities in obtaining optimal doses for synergistic effects. We propose to potentiate the synergy by enabling cis activation of IL-2 agonism and blockade of PD-1/PDL-2 binding with a single anti-PD1-IL2 to generate Bifunctional SuperKines for ImmunoTherapy (BiSKIT). Both human (h) and mouse (m) versions of anti-PD1 are fused with MDNA109FEAA, an IL-2 superkine with enhanced affinity for IL-2Rβ (CD122) and no binding to IL-2Rα (CD25).

Experimental Procedure: Studies included binding analyses with Biacore/SPR, signaling analyses in an IL-2 cell reporter assay, cell proliferation assay, in vitro pSTAT5 signaling in human PBMCs, PD1/PDL-1 cell reporter assay and efficacy in syngeneic tumor models.

Summary of Data: Anti-hPD1-MDNA109FEAA and surrogate anti-mPD1-MDNA109FEAA were tested in both in vitro and in vivo assays. Both constructs demonstrated selective binding to either hPD1 or mPD1 as expected, and the MDNA109FEAA motif retained its receptor selectivity (enhanced affinity to CD122 and no binding to CD25). In both HEK-Blue IL-2 reporter and CTLL-2 proliferation bioassays, anti-PD1-MDNA109FEAA exhibited similar agonistic effect as MDNA109FEAA-Fc, demonstrating that potency towards the IL-2 axis was not diminished. This was further confirmed with pSTAT5 signaling in primary human PBMCs where anti-PD1-MDNA109FEAA showed enhanced activation of CD8 T-cells and NK cells with reduced activity on Treg when compared to rhIL-2. Both constructs were also effective at inhibiting PD1/PDL-1 pathway in respective human and mouse specific cell-based reporter assays. In a CT26 syngeneic colon tumor model, anti-mPD1-MDNA109FEAA exhibited superior efficacy over monotherapy and combination with anti-mPD1 and MDNA109FEAA-Fc when administered at equal molar dosage and same schedule.

Conclusion: We have engineered anti-PD1-MDNA109FEAA to enable cis engagement with IL-2 receptor and PD1 as an approach to activate immune cells while reducing their exhaustion. Anti-mPD1-MDNA109FEAA achieved potent efficacy in a CT26 colon tumor model and is currently being evaluated in additional syngeneic tumor models with varying responsiveness to immune checkpoint blockade.

Citation Format: Minh D. To, Manjunatha Ankathatti Munegowda, Rosemina Merchant, Aanchal Sharma. An ‘anti-PD1-IL2 beta-only super-agonist’ displays potent anti-tumor efficacy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5532.
Abstract 5550: QLS31904: An anti-DLL3/CD3 bispecific antibody for T cell immunotherapy of small cell lung cancer
Authors: Yang Liuqing , Li Ruimei , Jiang Jiahua , Qian Hongliang , Ge Hongxiu , Lu Ting , Fang Yan , Liu Jun , Zhao Shuyong
Published: June 2022

Background: More than 150,000 patients are diagnosed with small cell lung cancer (SCLC) each year in the world. Standard of care for patients at different stages of SCLC includes radiation, etoposide, cisplatin/carboplatin, and atezolizumab. However, relapse is quite common. Therefore, there is a strong need to develop a more effective treatment. Delta-like ligand 3 (DLL3) is a target selectively expressed in SCLC rather than normal tissue. We have developed an anti-DLL3/CD3 bispecific antibody QLS31904 that is engineered to direct and activate T cells through CD3 to kill SCLC cells expressing DLL3.

Methods: Tumor cell killing, T cell activation, and cytokine production of QLS31904 in a concentration range were assessed in the presence of SCLC cells and human T cells in vitro. Antitumor activity of QLS31904 was evaluated in SCLC xenograft models, in which NCG mice were co-implanted with SHP-77 and human T cells first, then treated with intraperitoneal QLS31904 or control IgG1AA isotype. The in vivo tumor growth inhibition was assessed. Pharmacokinetic analyses were performed in non-human primates (NHP). The cross-reactivity of QLS31904 with NHP DLL3 and CD3 were established, and the tolerability was also evaluated in NHP model.

Results: QLS31904 displayed the anticipated bispecific binding properties and mediated potent and selective lysis of DLL3-positive cancer cell lines through recruitment and activation of human T cells. No cytokine activation was observed with human peripheral blood mononuclear cell alone. In xenograft models, QLS31904 induced significant tumor growth inhibition at doses as low as 20 µg/kg, including complete regression of established tumors. QLS31904 exhibits about 100 hours of T1/2 in NHP, supporting feasibility of weekly dosing. QLS31904 was well tolerated with no related adverse findings up to the highest tested dose (five weekly doses of 10mg/kg) in NHP.

Conclusions: QLS31904 has an impressive efficacy and safety profile in the preclinical studies, making it a potentially viable option for DLL3-expressing SCLC in the clinical setting.

Citation Format: Liuqing Yang, Ruimei Li, Jiahua Jiang, Hongliang Qian, Hongxiu Ge, Ting Lu, Yan Fang, Jun Liu, Shuyong Zhao. QLS31904: An anti-DLL3/CD3 bispecific antibody for T cell immunotherapy of small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5550.
Abstract 5559: Development of a novel PD-L1xCD40 bispecific antibody with excellent efficacy and safety profile for cancer therapy
Authors: luo Haishan , Meng Zihong , Rong Jimmy , Wang Yuandong , Yang Yunxing , Chen Fei , Wang Youhong , Chen Victor , Deng Gang , He Yebo , Gan Xin , Rong Yiping
Published: June 2022

CD40 agonistic antibody is emerging as a front-line immunotherapy for patients with cancer. However, despite its remarkable curative effects, serious immune-related adverse reactions limit the clinical application of CD40 agonists. Crosslinking-dependent activation is one of the promising approaches to diminish the adverse effects of CD40 agonists. Agonistic CD40 stimulation on antigen presenting cells (APCs) not only promotes activation of T cells, but also results in PD-L1 upregulation on APCs, thus creating a negative feedback loop to dampen the anti-tumor immune response. Therefore, we hypothesized that the combination of CD40 agonist and blockade of PD-1/PD-L1 interaction may further induce a synergistic effect on T cell activation and anti-tumor activity. Here, we report a novel PD-L1xCD40 Bispecific Antibody (BsAb) designed to have synergistic curative effects as well as improved safety profiles. The PD-L1/CD40 BsAb was generated using the Harbour HBICE࣪® platform. It exhibited superior binding activities to both PD-L1 and CD40, respectively. CD40 activation was fully relied on the PD-L1 crosslinking. The PD-L1xCD40 BsAb showed potent tumor growth inhibition in preclinical efficacy models. Meanwhile, it showed much improved safety profile compared to a reference anti-CD40 agonistic antibody in the preclinical tox models. Overall, the PD-L1xCD40 BsAb demonstrated remarkable in vivo anti-tumor efficacy and improved safety profiles in preclinical models, which presents it as a promising candidate for the development of next-generation cancer immune therapeutics.

Citation Format: Haishan luo, Zihong Meng, Jimmy Rong, Yuandong Wang, Yunxing Yang, Fei Chen, Youhong Wang, Victor Chen, Gang Deng, Yebo He, Xin Gan, Yiping Rong. Development of a novel PD-L1xCD40 bispecific antibody with excellent efficacy and safety profile for cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5559.
Abstract 5697: Targeting PARP inhibitor resistance with Polθ inhibitors
Authors: Zatreanu Diana , Robinson Helen , Alkhatib Omar , Boursier Marie , Finch Harry , Geo Lerin , Grande Diego , Grinkevich Vera , Heald Robert , Langdon Sophie , Majithiya Jayesh , McWhirter Claire , Martin Niall , Moore Shaun , Neves Joana , Rajendra Eeson , Ranzani Marco , Schaedler Theresia , Stockley Martin , Wiggins Kimberley , Brough Rachel , Sridhar Sandhya , Gulati Aditi , Shao Nan , Badder Luted , Novo Daniela , Knight Eleanor , Marlow Rebecca , Haider Syed , Callen Elsa , Hewitt Graeme , Schimmel Joost , Prevo Remko , Alli Christina , Ferdinand Amanda , Bell Cameron , Blencowe Peter , Bot Chris , Calder Mathew , Charles Mark , Curry Jayne , Ekwuru Tennyson , Nussenzweig Andre , Tijsterman Marcel , Tutt Andrew N. , Boulton Simon , Higgins Geoff , Pettitt Stephen J. , Smith Graeme C. , Lord Christopher J.
Published: June 2022

To target DNA repair vulnerabilities in cancer, we discovered nanomolar potent, selective, low molecular weight (MW), allosteric inhibitors of the polymerase function of DNA polymerase Polθ, including ART558. ART558 inhibits the major Polθ-mediated DNA repair process, Theta-Mediated End Joining (TMEJ), without targeting Non-Homologous End Joining. Moreover, we show that exposure to ART558 can elicit DNA damage and synthetic lethality in BRCA1- or BRCA2-mutant tumor cells and enhances the effects of a PARP inhibitor. Genetic perturbation screening revealed that defects in the 53BP1/Shieldin complex, which are a cause of PARP inhibitor resistance, result in in vitro and in vivo sensitivity to Polθ polymerase inhibitors. Mechanistically, ART558 increases biomarkers of single-stranded DNA and synthetic lethality in 53BP1-defective cells. The inhibition of DNA nucleases that promote end-resection, such as Exo1 or Blm-Dna2 reversed these effects, implicating these in the synthetic lethal mechanism-of-action. Taken together, these observations describe a drug class that elicits BRCA-gene synthetic lethality and PARP inhibitor synergy, as well as targeting a biomarker-defined mechanism of PARPi-resistance.

Citation Format: Diana Zatreanu, Helen Robinson, Omar Alkhatib, Marie Boursier, Harry Finch, Lerin Geo, Diego Grande, Vera Grinkevich, Robert Heald, Sophie Langdon, Jayesh Majithiya, Claire McWhirter, Niall Martin, Shaun Moore, Joana Neves, Eeson Rajendra, Marco Ranzani, Theresia Schaedler, Martin Stockley, Kimberley Wiggins, Rachel Brough, Sandhya Sridhar, Aditi Gulati, Nan Shao, Luted Badder, Daniela Novo, Eleanor Knight, Rebecca Marlow, Syed Haider, Elsa Callen, Graeme Hewitt, Joost Schimmel, Remko Prevo, Christina Alli, Amanda Ferdinand, Cameron Bell, Peter Blencowe, Chris Bot, Mathew Calder, Mark Charles, Jayne Curry, Tennyson Ekwuru, Andre Nussenzweig, Marcel Tijsterman, Andrew N. Tutt, Simon Boulton, Geoff Higgins, Stephen J. Pettitt, Graeme C. Smith, Christopher J. Lord. Targeting PARP inhibitor resistance with Polθ inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5697.
Abstract 587: ACLX-002, a novel CD123-targeted universal CAR-T cell therapy for relapsed or refractory acute myeloid leukemia that can be activated and silenced in vivo with soluble protein adapters in a dose dependent manner
Authors: Edwards Justin P. , Zaritskaya Liubov , Swers Jeff , Ng Sinnie , Mu Jenny , Witter Alexandra , Cheung Laurene , McCullough Samantha , LaFleur David , Hilbert David , Tice David
Published: June 2022

Genetically engineered T cells have demonstrated great promise in the treatment of hematologic malignancies including Acute Lymphocytic Leukemia, Non-Hodgkin’s lymphoma and Multiple Myeloma. However, those successes are often associated with serious dose-limiting adverse events including cytokine release syndrome (CRS) and immune effector-cell associated neurotoxicity syndrome (ICANS), as well as frequent tumor relapse. Specifically, in AML, there is a need for controllable and adaptable cellular therapeutics that can address intra- and inter- patient disease heterogeneity and off-target antigen expression that contribute to these dose-limiting adverse events. Herein, we report a novel T cell therapy, ACLX-002, comprised of a soluble tumor targeting protein (SPRX002) that specifically binds CD123-expressing AML cells and targets those cells for destruction by ex vivo transduced T cells known as Antigen Receptor Complex (ARC) T cells. Functional in vitro studies of ARC-T cells produced from healthy subjects demonstrated SPRX002 dose-dependent cytokine production, T cell proliferation, and cytotoxic activity of co-cultured CD123-expressing cancer cell lines. The cytolytic machinery of the ARC-T cells is only activated when the tri-complex of the ARC-T cell, SPRX002, and CD123-expressing cell is fully formed. ACLX-002 also demonstrated dose- and schedule-dependent in vivo efficacy in both the MOLM14 and MV4-11 disseminated CD123-expressing tumor models in NSG immunocompromised mice, as well as multiple AML patient derived xenograft models. A single dose of 5 x 106 ARC-T cells together with SPRX002 doses of 0.3 mg/kg daily were able to eliminate measurable tumor burden that was similar to a CD123-directed traditional CAR-T that uses the same CD123-binding domain as SPRX002. Furthermore, the in vivo efficacy was not limited by the expression of CD123 on donor T cells, an attribute that is induced by in vitro activation and expansion of ARC-T cells. The data support the paradigm that ARC-T cells can be activated and silenced by controlling the dose and schedule of administered SPRX002, which may improve the safety and effectiveness of T cell therapy.

Citation Format: Justin P. Edwards, Liubov Zaritskaya, Jeff Swers, Sinnie Ng, Jenny Mu, Alexandra Witter, Laurene Cheung, Samantha McCullough, David LaFleur, David Hilbert, David Tice. ACLX-002, a novel CD123-targeted universal CAR-T cell therapy for relapsed or refractory acute myeloid leukemia that can be activated and silenced in vivo with soluble protein adapters in a dose dependent manner [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 587.
Abstract 5881: The efficacy of a technology-based support program on survivorship experience: Asian American breast cancer survivors
Authors: Im Eun-Ok , Chee Wonshik , Paul Sudeshna ,   TICAA Research Team
Published: June 2022

Purpose: A technology-based intervention without face-to-face interactions could work better for many women from cultures where breast cancer is still a stigmatizing experience. Also, a technology-based intervention could meet the necessity of information and support without time and/or geographical restrictions. The purpose of this study was to test the efficacy of a technology-based support program for Asian American breast cancer survivors (TICAA) in enhancing the women’s breast cancer survivorship experience.

Methods: This was a randomized controlled intervention with repeated measures (pretest [Time Point 0; T0], post 1-month [T1], and post 3-months [T2]). A total of 199 Asian American breast cancer survivors were recruited/retained through online and offline settings and included in the data analysis. While the intervention group was requested to use the TICAA and the American Cancer Society (ACS) website on breast cancer for 3 months, the control group was requested to use the ACS website for 3 months. Multiple instruments were used: the Cancer Behavior Inventory (CBI), the Personal Resource Questionnaire (PRQ), the Perceived Isolation Scale (PIS), the Support Care Needs Survey-34 Short Form (SCNS), the Memorial Symptom Assessment Scale-Short Form (MSAS), and the Functional Assessment of Cancer Therapy Scale-Breast Cancer (FACT-B). The data were analyzed using growth curve models, fitted within the framework of mixed effects modeling, and mediation analyses.

Results: Within the mixed-effect model, there were significant decreases in the needs for help scores (SCNS scores) from T0 to T2 (β = -3.0236, p = .005) and the symptom distress scores (MSAS scores) from T0 to T1 and T2 (β = -2.8131, p < .001; β = -3.0356, p < .001). Yet, the change over time in the quality-of-life scores (FACT-B scores) was not statistically significant although the fixed effect of time was significant (β = -1.0476, p = .004). All the theory-based mediators (attitudes, self-efficacy, perceived barriers, and social influences) had statistically significant mediating impact on all outcome variables (needs for help, symptom distress, and quality-of-life; all p < .001), but there were differences in the mediating effects among different individual outcome variables.

Conclusions: The findings supported the efficacy of the technology-based program in reducing the need for help and symptoms of Asian American breast cancer survivors. Further studies are needed on the differences in the effects of theory-based mediators on different outcome variables.

Citation Format: Eun-Ok Im, Wonshik Chee, Sudeshna Paul, TICAA Research Team. The efficacy of a technology-based support program on survivorship experience: Asian American breast cancer survivors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5881.
Abstract 5979: Evaluating therapeutic strategies against lung cancer using multispectral optoacoustic tomography
Authors: Wang Tianyuan , Wanniarachchi Hashini , Arango Lorena , Zhang Li , Liu Li
Published: June 2022

Tumor hypoxia is an important feature of the microenvironment in various types of tumors. The leaky and compressed vessels in the tumor lead to impaired blood flow and oxygen delivery and cause hypoxia. Significant tumor hypoxia exists in non-small cell lung cancer (NSCLC) and is closely associated with radiation-resistant and poor survival. Recently, we have identified a new class of antitumor drugs, cyclopamine tartrate (CycT) and heme-sequestering peptide 2 (HeSP2). They are heme-targeting agents that effectively inhibit NSCLC oxidative metabolism, oxygen consumption, and ATP generation, which lead to the suppression of tumor progression. By using multispectral optoacoustic tomography (MSOT), an advanced noninvasive imaging modality that monitors the changes in blood oxygen saturation and vascular function in real-time, we observed the effect of heme-targeting agents in normalizing tumor vasculature and alleviating tumor hypoxia. To investigate the impact of heme-targeting agent on mediating the lung cancer radiotherapy response, we characterized the vasculature and antigenic profile of NSCLC growth during the treatment of CycT in combination with radiotherapy by performing MSOT, together with other imaging approaches including bioluminescence imaging (BLI) and immunohistochemistry (IHC). We demonstrated that the heme-targeting agents increased tumor radiation sensitivity by regulating tumor vasculature and oxygenation. Our novel combination therapy can be a powerful strategy in improving the treatment of lung cancer and potentially other radiation-resistant cancers.

Citation Format: Tianyuan Wang, Hashini Wanniarachchi, Lorena Arango, Li Zhang, Li Liu. Evaluating therapeutic strategies against lung cancer using multispectral optoacoustic tomography [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5979.
Abstract 5988: Intra-carotid artery brain metastasis models for evaluation of lung and breast cancer drugs
Authors: Wang Fuyang , Yang Yuying , Zhao Haoxia , Dai Chunlei , Han Bingrui , Zhang Mengyao , Xiong Xia , Zhang Chao , Tang Xuzhen , Ji Qunsheng
Published: June 2022

Non-CNS metastatic brain cancer is about 10 times more common than CNS cancer. Lung cancer and breast cancer account for most brain metastasis. KRAS-mutant NSCLC compose a third lung adenocarcinoma, among which 17% to 55% will develop brain metastases. Likewise, more than a third HER2-positive breast cancer will develop brain metastasis. Among existing animal models, the ectopic injection in brain cannot reflect the mechanism of tumor invasion and metastasis, while tail vein and intra-cardiac injection usually produces extra-cerebral metastatic disease and the animals have to be sacrificed before brain metastases appeared. Here we developed several metastasis models injected by intra-carotid artery (ICA) to address these problems. Two KRAS-mutant lung cancer cells and two HER2-positive breast cancer cells selected by their metastasize potential were luciferase labeled for in vivo imaging and drug evaluation. The tumor pathology in brain was evaluated by H&E staining. Compared to ectopic models, these ICA models have natural dissemination and progression in the brain, which recapitulate the different pattern of metastatic lesions in the clinic. With synchronized growth rate and little variation compared to tail vein and intra-cardiac models, these tumor growth kinetics allow for more appropriate animal efficacy study. Both KRAS-sensitive and insensitive models have been developed and validated for the next generation drug development; HER-2 positive breast models with ICA method also showed a good discriminative power on anti-HER2 modalities including antibodies, small molecules and ADC. Meanwhile, ICA models with human immune cell reconstitution were also developed to support immune therapies in pre-clinical stage. In summary, we have developed a panel of intra-carotid artery brain metastatic models for drug evaluation, and demonstrated their superiority in growth kinetics, brain microenvironment, drug selectivity and immune component. These models can be a promising predictive tool for novel drug/modality discovery to target brain metastasis.

Citation Format: Fuyang Wang, Yuying Yang, Haoxia Zhao, Chunlei Dai, Bingrui Han, Mengyao Zhang, Xia Xiong, Chao Zhang, Xuzhen Tang, Qunsheng Ji. Intra-carotid artery brain metastasis models for evaluation of lung and breast cancer drugs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5988.
Abstract 6051: Longitudinal single-cell sequencing of high-risk neuroblastoma tumors reveals intrinsic and extrinsic mechanisms of therapy resistance
Authors: Uzun Yasin , Grossmann Liron D. , Chen Chia-Hui , Thadi Anusha , Wu Chi-Yun , Gao Peng , Diep Dinh , Surrey Lea , Martinez Daniel , Patel Tasleema , Qiu Qi , Johnson Sarah , Yu Wenbao , Drabings Shane , Chen Changya , Hu Yuxuan , Chen Gregory , Oldridge Derek A. , Zhang Kun , Wu Hao , Bernt Kathrin , Zhang Nancy , Maris John M. , Tan Kai
Published: June 2022

Neuroblastoma is a childhood cancer originating from embryonic neuronal progenitor cells and is the most common cancer diagnosed in infants. Although the majority of patients respond to initial chemotherapy, most high-risk patients suffer relapse due to therapy resistance.

Here, we generated single-cell transcriptome and bulk whole-genome sequencing data of diagnosis and post-therapy samples from 23 patients diagnosed with high-risk neuroblastoma. We found two distinct adrenergic populations in the patient samples. Most tumor cells showed a mature sympathoblast phenotype whereas a small fraction was in an undifferentiated state with activation of protein translation, unfolded protein, and oxidative phosphorylation pathways. We found that therapy-resistant tumor cells in these two subpopulations had distinct characteristics. The more mature resistant subpopulation upregulated genes associated with epithelial-to-mesenchymal transition, including TWIST1, PLCB1 and CD276. The undifferentiated resistant subpopulation upregulated Ras signal transduction and TP53 pathway genes.

Analysis of the immune microenvironment revealed that most tumor-associated macrophages became more immunosuppressive post-therapy via multiple newly gained signaling interactions including the complement signaling pathway. We discovered a limited infiltration of T lymphocytes in the tumor microenvironment, and chemotherapy induced an effector state with upregulated mTOR signaling and metabolism. Overall, our study revealed subpopulations of tumor cells in neuroblastoma that responded differently to induction chemotherapy. Our findings uncovered distinct molecular signatures of the resistant cells and their interactions with the immune microenvironment, paving the way for developing novel therapies for high-risk neuroblastoma.

Citation Format: Yasin Uzun, Liron D. Grossmann, Chia-Hui Chen, Anusha Thadi, Chi-Yun Wu, Peng Gao, Dinh Diep, Lea Surrey, Daniel Martinez, Tasleema Patel, Qi Qiu, Sarah Johnson, Wenbao Yu, Shane Drabings, Changya Chen, Yuxuan Hu, Gregory Chen, Derek A. Oldridge, Kun Zhang, Hao Wu, Kathrin Bernt, Nancy Zhang, John M. Maris, Kai Tan. Longitudinal single-cell sequencing of high-risk neuroblastoma tumors reveals intrinsic and extrinsic mechanisms of therapy resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6051.
Abstract 6087: Evaluation of intra-tumor heterogeneity by single-cell copy number profiling of circulating multiple myeloma cells using Shannon index
Authors: Ferrarini Alberto , Forcato Claudio , Monaco Valentina del , Terracciano Mario , Garonzi Marianna , Pellicano Massimiliano , Buson Genny , Fontana Francesca , Tononi Paola , Manaresi Nicolo
Published: June 2022

Tumors develop through an evolutionary process in which genomic lesions accumulate in different coexisting subclones leading to intra-tumor heterogeneity (ITH). ITH allows the tumor to adapt to microenvironments and to resist therapies and has been shown to be a prognostic marker. In this work we show the characterization of ITH in Multiple Myeloma (MM) at single cell resolution from a pool of Circulating Multiple Myeloma Cells (CMMCs) and we study the relationship between number of sampled cells and diversity detected.

Peripheral blood was collected from 3 patients affected by MM. Using CellSearch® system, blood was enriched for target CMMCs, with immuno-magnetic selection of CD138+, and immunofluorescently stained cells, positive for CD38-PE, DAPI and negative for CD45/CD19-APC, were enumerated. The samples were then analyzed and CMMCs were isolated with DEPArray platform. The DNA of each single cell (n=261) was amplified with Ampli1™ WGA kit and the WGA output was used for genome-wide single cell copy number alteration analysis with Ampli1 LowPass kit.

Copy number profiling revealed from 2 to 3 coexisting subclones per patient, with minor clones size going from 6.5% to 33%. ITH was assessed by determining the Shannon index, which showed a range of values from 0.44 (low diversity) to 1.01 (high diversity) depending on source sample. We evaluated the effect of number of cells/profiles on Shannon index by bootstrapping (200 times) a number of copy number profiles ranging from 2 to the total number of profiles available per patient. Results showed that the mean Shannon index almost reaches a plateau at 15-20 cells/profiles in the case of lower diversity samples and at about 30 cells/profiles for the highest diversity sample. Moreover, as few as 10 cells allowed to differentiate low vs. high diversity samples.

Cell enrichment and sorting by CellSearch and DEPArray platforms allow to quantify ITH at diversity level with high precision even from a limited number of initial single-cells and provide a promising method to objectively measure heterogeneity, as required in order to study its clinical implications.

Citation Format: Alberto Ferrarini, Claudio Forcato, Valentina del Monaco, Mario Terracciano, Marianna Garonzi, Massimiliano Pellicano, Genny Buson, Francesca Fontana, Paola Tononi, Nicolo Manaresi. Evaluation of intra-tumor heterogeneity by single-cell copy number profiling of circulating multiple myeloma cells using Shannon index [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6087.
Abstract 6097: Ferroptosis resistance in epithelial populations of triple negative breast cancer is mediated by laminin 332 and integrin beta4
Authors: Karner Emmet R. , Goel Hira Lal , Mercurio Arthur M.
Published: June 2022

Ferroptosis is an iron dependent form of regulatory cell death caused by the accumulation of lipid peroxides that can be exploited as a therapeutic for carcinomas and possibly other cancers. However, some populations of carcinoma cells resist ferroptosis, especially those with an epithelial phenotype but the mechanisms that promote resistance are poorly understood. We discovered that ferroptosis resistance in triple-negative breast cancer cells (TNBC) with an epithelial phenotype is dependent on expression of the integrin β4 and its ligand laminin 332 (LM332). LM332 and the integrin β4 maintain an epithelial cell state by downregulation of the EMT factor ZEB1. We further demonstrate that repression of ZEB1 by integrin β4 and LM332 promotes YAP/TAZ-mediated transcription of the C2 subunit of LM332, activating a positive feedback loop. Lastly, we observed that disrupting the β4/LM332 signaling axis sensitizes epithelial TNBC cells to ferroptosis by inducing ZEB1 expression. Our findings reveal a novel role for the extracellular matrix in promoting ferroptosis resistance by sustaining an epithelial phenotype and they suggest potential therapeutic strategies for overcoming this resistance.

Citation Format: Emmet R. Karner, Hira Lal Goel, Arthur M. Mercurio. Ferroptosis resistance in epithelial populations of triple negative breast cancer is mediated by laminin 332 and integrin beta4 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6097.
Abstract 6137: Tumor-derived endothelin-1 recruits and activates fibroblasts to support tumor aggressiveness
Authors: Rio Danila Del , Caprara Valentina , Masi Ilenia , Spadaro Francesca , Giannitelli Sara , Rainer Alberto , Bagnato Anna , Rosanò Laura
Published: June 2022

The recruitment of fibroblasts to tumor and their activation to cancer-associated fibroblast (CAF) is an exploited strategy used by tumor to guide matrix remodeling, supporting cancer invasion and metastatic disease. CAFs are the major effectors of extracellular matrix (ECM) remodelling through secretion of collagens, cross-linking enzymes and proteases, and in turn engagement of integrins. Although several tumor cell-secreted factors have been identified in generating tumor-promoting stroma, there has been a great interest to define factors and associated molecular mechanisms involved in promoting CAF premetastatic function. Tumor-related endothelin-1 (ET-1) has an important role in the crosstalk between cancer and stromal cells supporting serous ovarian cancer (SOC) progression. By binding its receptors (ETAR and ETBR), ET-1 favours the recruitment of the protein β-arrestin-1 and the generation of signaling complexes regulating also cytoskeleton reorganization, thereby fine-tuning SOC cell invasion and metastasis. However, how the integration of ET-1 receptors might “educate” stroma to become permissive for invasion, supplying an altered ECM governing metastasis needs to be investigated. In this study, we identified endothelin-1 as a key factor in priming CAF conversion in SOC. We demonstrate that primary ovarian fibroblasts express ETA and ETB, along with β-arrestin1, and secrete ET-1; b) the autocrine and cancer-secreted ET-1 promotes fibroblast proliferation, and an increase in the expression of CAF markers (αSMA, vimentin and FAP), secretion of proinflammatory cytokines, collagen contraction ability and cell motility. Moreover, ET-1 induces activation of b1 integrin and downstream signaling. Mechanistically, ET1R/intb1 signaling allows ECM remodeling, either by promoting and invasive protrusion formation, invadosome, and collagen secretion. Importantly, therapeutical inhibition of ET-1 receptors with Ambrisentan and intβ1 with ATN-116 markedly reduced the ability of SOC cells to adhere in the intraperitoneal organs and to metastasize. Our findings support a model in which ET-1 educates the stromal fibroblasts to become permissive for tumor invasion and demonstrate for the time the significance of an ET-1-dependent integrin signaling in this process.

Citation Format: Danila Del Rio, Valentina Caprara, Ilenia Masi, Francesca Spadaro, Sara Giannitelli, Alberto Rainer, Anna Bagnato, Laura Rosanò. Tumor-derived endothelin-1 recruits and activates fibroblasts to support tumor aggressiveness [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6137.
Abstract 6306: Genomic analysis markers of stemness in oral cancer progression
Authors: Unger Gretchen , Wuertz Beverly G. , Gaffney Patrick , Ondrey Frank G.
Published: June 2022

Introduction: In clinical cancer treatment there is an continuing effort to identify limited gene sets from high dimensional data to predict risk, treatment response, etc. Oral carcinogenesis is a multistep process of mutations, disordered growth, and the development of invasion. In the oral cancer milieu, inflammation and stemness has been demonstrated to be cooperative during carcinogenesis. We hypothesized that differences in inflammation, stemness, and EMT might distinguish lesion and adjacent normal mucosa in our RNA Seq data on preneoplastic lesions. Once candidate genes are identified they might be applied to other oral mucosal specimens to identify risk for malignancy development.

Materials and Methods: We collected lesion and adjacent normal mucosal tissues from 16 subjects with high risk leukoplakia and early oral cancer (T1N0 and T2N0). Sequencing libraries were generated using the Illumina Truseq Stranded mRNA library prep kit. Sequencing was performed on an Illumina Hiseq 3000 instrument with paired-end 75bp read and sequenced to an overall depth of 30 million reads per sample.

Results: After developing a biospecimen collection procedure that functions in the senior author’s clinic flow, we were able to isolate sequenceable total RNA from 100% of the specimens we collected. We identified several hundred differentially expressed genes between lesion and adjacent normal. With regard to stemness, inflammation, and EMT we multiple candidate genes that could be incorporated into a limited gene set for further studies. Aldehyde dehydrogenase family members were of particular interest in the stemness candidate genes, for example. ALD 1A1 and ALD 3B1, were significantly upregulated in lesion versus normal (P< 0.02).

Conclusions: We conclude there are differentially expressed inflammatory, EMT, and stemness in oral preneoplastic specimens. We have identified multiple candidate genes that could be assembled into a limited gene panel (12-24 genes) for further studies of oral cancer risk in mucosal lesions

Citation Format: Gretchen Unger, Beverly G. Wuertz, Patrick Gaffney, Frank G. Ondrey. Genomic analysis markers of stemness in oral cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6306.