Solid Tumor Patients Research Articles

Elective Checkpoint Inhibitor Discontinuation in Metastatic Solid Tumor Patients: A Case Series.

Checkpoint inhibitor (CKI) therapy has markedly altered the survival of patients with many solid tumors. It appears clear that 10-40% of patients with a number of metastatic cancers can achieve lengthy remissions following CKI therapy. The optimal duration of treatment or whether treatment can ever be safely stopped is still controversial. Based on melanoma-derived data, we tested whether CKI treatment could safely be discontinued in patients with other solid tumors. A retrospective analysis was performed in adults with metastatic solid tumors treated with CKI-based therapy. Patients with solid tumors who achieved complete remission on 2 sequential scans at least 3 months apart during ongoing treatment were identified from our computerized patient database. Patient data was analyzed for patient characteristics, as well as progression-free and overall survival. A total of 69 non-melanoma solid tumor patients were treated with CKI-based regimens in our clinic and 14 achieved complete remission (20.3%). Five patients were female (35.7%) and the remaining nine were male (64.3%). A 100% progression-free survival was observed for these patients. The median duration of complete remission was over 20 months from the time of elective treatment discontinuation. Median overall survival was not reached in this cohort. One patient died of no cancer-related causes. Based on this retrospective case series, elective treatment discontinuation in patients who achieved complete remission appears feasible. All patients remained in a durable complete remission after treatment discontinuation. We hypothesize that appropriate selection of patients for early treatment discontinuation may decrease their economic burden related to ongoing treatment, limit potential toxicity, and improve quality of life.

The efficient circulating immunoscore predicts prognosis of patients with advanced gastrointestinal cancer

BackgroundImmunoscore from tumor tissues was initially established to evaluate the prognosis of solid tumor patients. However, the feasibility of circulating immune score (cIS) for the prognosis of advanced gastrointestinal cancers (AGC) has not been reported.Material and methodsPeripheral venous blood was collected from 64 untreated AGC patients. We utilized flow cytometry to determine several immune cell subpopulations, including CD8+ and CD4+ T cells, NK cells, and CD4 + CD25 + CD127low Tregs. The circulating immune score 1 (cIS1) was assessed according to the proportions of CD4+, CD8+ T cells, and NK cell, whereas circulating immune score 2 (cIS2) was derived from the proportions of CD4+, CD8+ T cell, and CD4 + CD25 + CD127low Tregs. The prognostic role of cIS for progression-free survival (PFS) and overall survival (OS) was analyzed using Kaplan–Meier curves and Cox multivariate models. Receiver operating characteristic (ROC) curves were depicted to compare the prognostic values of cIS1 and cIS2.ResultsAGC patients with high cIS1(≥ 2) and cIS2(≥ 2) had significantly longer PFS (cIS1: median PFS, 11 vs. 6.7 months, P = 0.001; cIS2: 12 vs. 5.8 months, P < 0.0001) and OS (cIS1: median OS, 12 vs. 7.9 months, P = 0.0004; cIS2: 12.8 vs. 7.4 months, P < 0.0001) than those with low cIS1 and low cIS2. The areas under ROC curves (AUROCs) of cIS1 and cIS2 for OS were 0.526 (95% confidence interval; 95% CI 0.326–0.726) and 0.603 (95% CI 0.427–0.779, P = 0.332), whereas AUROC of cIS2 for PFS was larger than that of cIS1 0.735 (95% CI 0.609–0.837) vs 0.625 (95% CI 0.495–0.743) (P = 0.04)).ConclusionThe cIS can be applied to predict the prognosis of untreated AGC patients. Compared with cIS1, cIS2 displayed superior prognostic value for PFS prediction.

Analysis of Extended Length of Stay in a Comprehensive Cancer Center

Background: The management of cancer patients is complex and usually requires hospital admission in order to diagnose and treat complications related to cancer or its therapies. Solid tumor patients have a mean length of stay (LOS) of about 6 to 9 days. A large portion of the cancer patients has extended length of stay more than 10 days. Long hospital stays utilize more resources and is more expensive. Methods: We aimed to assess characteristics of patients with an extended length of stay at UT MD Anderson OncoHospitalist service in 2019 to identify focus areas that can help decrease extended LOS. Results: 172 out of 1449 patients with extended LOS were analyzed. This study has a 95% confidence interval of finding the true value within (±7%) of the real value. The results showed that 84% of prolonged admissions were due to medical issues and 18% of the patients were either discharged to hospice or died during their hospital stay. Further breakdown of the medical reasons for extended LOS indicated less 4% of these patients received inpatient radiation therapy; around 3% had complications from immunotherapy; 2% patients had issues with pain control and 7% of these patients had issues with placement. Overall, 50% of the patients who had issues with placement had Medicare and 25% had no health insurance. The study was unable to determine any association between the distances of primary residence from hospital to extended LOS. Conclusion: The study was able to identify focus areas such as early Goals of Care (GOC) in end-of-life care, protocolization of immunotherapy adverse reaction therapy, and criteria-based approach to earlier radiation oncology consultation for palliative radiation therapy. All of which will help decrease the extended LOS at places like UT MD Anderson and others.

A nationwide study on characteristics and outcome of cancer patients with sepsis requiring intensive care

Background Sepsis is the leading cause of admission to the intensive care unit (ICU) for cancer patients and survival rates have historically been low. The aims of this nationwide cohort study were to describe the characteristics and outcomes of cancer patients admitted to the ICU with sepsis compared with other sepsis patients requiring ICU admission. Material and methods This was a retrospective, observational study. All adult admissions to Icelandic ICUs during years 2006, 2008, 2010, 2012, 2014 and 2016 were screened for severe sepsis or septic shock by ACCP/SCCM criteria. Clinical characteristics and outcomes of sepsis patients with cancer were compared to those without cancer. Results In the study period, 235 of 971 (24%) patients admitted to Icelandic ICUs because of sepsis had cancer, most often a solid tumour (100), followed by metastatic tumours (69) and haematological malignancies (66). Infections were more often hospital-acquired in cancer patients (52%) than other sepsis patients (18%, p < 0.001) and sites of infections differed, with abdominal infections being most common in patients with solid and metastatic tumours but lungs and bloodstream infections in haematological malignancies. The length of stay in the ICU was shorter for sepsis patients with metastatic disease than other sepsis patients (2 vs. 4 days, p < 0.001) and they were more likely to have treatment limitations (52 vs. 19%, p < 0.05). Median survival of patients with metastatic disease was 19 days from ICU admission. The 28-day mortality (25%) of solid tumour patients was comparable to that of sepsis patients without cancer (20%, p < 0.001). Conclusions Cancer is a common comorbidity in patients admitted to the ICU with sepsis. The clinical presentation and outcome differs between cancer types. Individuals with metastatic cancer were unlikely to receive prolonged invasive ICU care treatment. Comparable short-term outcome was found for patients with solid tumours and no cancer.

Genomic Features of Solid Tumor Patients Harboring ALK/ROS1/NTRK Gene Fusions.

The fusions of receptor tyrosine kinase (RTK) involving anaplastic lymphoma kinase (ALK), c-ros oncogene 1 (ROS1), and neurotrophic receptor tyrosine kinase (NTRK) represent the potential targets of therapeutic intervention for various types of solid tumors. Here, the genomic features of 180 Chinese solid tumor patients with ALK, ROS1, and NTRK fusions by next generation sequencing (NGS) were comprehensively characterized, and the data from 121 patients in Memorial Sloan Kettering Cancer Center (MSKCC) database were used to compare. We found that ALK, ROS1, and NTRK fusions were more common in younger female patients (p<0.001) and showed a higher expression of programmed death ligand 1 (PD-L1). The gene-intergenic fusion and the fusion with rare formation directions accounted for a certain proportion in all samples and 62 novel fusions were discovered. Alterations in TP53 and MUC16 were common in patients with RTK fusions. The mutational signatures of patients were mainly distributed in COSMIC signature 1, 2, 3, 15 and 30, while had a higher frequency in copy number variations (CNVs) of individual genes, such as IL-7R. In the MSKCC cohort, patients with fusions and CNVs showed shorter overall survival than those with only fusions. Furthermore, the differentially mutated genes between fusion-positive and -negative patients mainly concentrated on MAPK signaling and FOXO signaling pathways. These results may provide genomic information for the personalized clinical management of solid tumor patients with ALK, ROS1, and NTRK fusions in the era of precision medicine.

When targeted therapy for cancer leads to ICU admission. RETRO-TARGETICU multicentric study

To study prevalence of targeted therapy (TT)-related adverse events requiring ICU admission in solid tumor patients. Retrospective multicenter study from the Nine-i research group. Adult patients who received TT for solid tumor within 3 months prior to ICU admission were included. Patients admitted for TT-related adverse event were compared to those admitted for other reasons. In total, 140 patients, median age of 63 (52-69) years were included. Primary cancer site was mostly digestive (n=27, 19%), kidney (n=27, 19%), breast (n=24, 17%), and lung (n=20, 14%). Targeted therapy was anti-VEGF/VEGFR for 27% (n=38) patients, anti-EGFR for 22% (n=31) patients, anti-HER2 for 14% (n=20) patients and anti-BRAF for 9% (n=5) patients. ICU admission was related to TT adverse events for 30 (21%) patients. The most frequent complications were interstitial pneumonia (n=7), cardiac failure (n=5), anaphylaxis (n=4) and bleeding (n=4). At ICU admission, no significant difference was found between patients admitted for a TT-related adverse event and the other patients. One-month survival rate was higher in patients admitted for TT adverse event (OR=5.733 [2.031-16.182] P<0.001). Adverse events related to targeted therapy accounted for 20% of ICU admission in our population and carried a 16% one-month mortality. Outcome was associated with admission for TT related to adverse event, breast cancer and good performance status.

Abstract 1254: Actionable fusions detected by RNA-seq co-occur with PD-L1 expression and driver mutations in solid tumors patients

Abstract Background: Incorporating gene fusions into a comprehensive profile is critical not only because very effective therapies targeting oncogenic fusion proteins exist but they may also negate the response to therapy of an actionable SNV and InDel mutations. We examined the co-occurrence of actionable gene fusions detected by RNA-seq with actionable SNVs/InDels and with the immunotherapy response biomarker, PD-L1. Methods: In 2021, 5341 FFPE samples were analyzed by our clinical laboratory using a novel hybridization-based RNA sequencing assay. DNA (SNV/Indels) mutations were detected with a clinical grade NGS assay and PD-L1 protein expression was determined by IHC using an appropriate LDT or FDA approved assay. De-identified data were analyzed following an approved IRB protocol. Results: Of the 5341 patients tested for gene fusions only 0.7% were profiled with a comprehensive fusion detection panel for 250 clinically relevant fusion genes. Conversely, 67% of all patients were tested only for NTRK gene fusions. The prevalence of the following most relevant fusions was: ALK=3.16% (in particular EML4-ALK), NTRKs=0.85%, FGFR2=5.60%, RET=1.34% and ROS1=1.05%. Among NTRK fusions, NTRK3 was detected in 52.9% of the positive cases; in particular, dominant fusions are ETV6-NTRK3 (26%) and EML4-NTRK3 (20%). In the 163 fusion positive cases, 37 cases had available DNA mutation testing and PD-L1 expression testing. These patients presented 24 different actionable fusions, including ALK (2), FGFR1/2 (5), NTRK 1/2/3 (10), NRG1 (3), RET (3), and ROS1 (3) fusions. Interestingly, while 73% (27/37) of those tumors were PD-L1 positive, similar to the 75% found on the fusion negative samples, PD-L1 was positive in 88% (15/17) of the lung samples with pathogenic mutations from this subset. This was strikingly higher than the 45% (192/424) found in the fusion negative cohort. Finally, excluding TP53 mutations, NTRK fusions frequently co-occurred with RNF43, FBXW7, TERT promoter, and ARID1A pathogenic mutations. FGFR fusions co-occurred with BAP1, PBRM1, or KRAS mutations, which correlates with the fact that both FGFR2 fusions and swi/snf alterations are enriched in intrahepatic cholangiocarcinoma, where the FGFR fusions were detected. RET fusions co-occurred with ARID1A and TERT. ROS1 fusions co-occurred with SMARCA4 and KRAS pathogenic mutations simultaneously. NRG1 fusions co-occurred with ARID1A and FBXW7 mutations. EML4-ALK fusions co-occurred with FGFR2 and KMT2D variants of unknown significance. Conclusion: Actionable cancer-driving gene fusions were detected by RNA-sequencing and co-occurred with other biomarkers that can guide the selection of therapies, such as immune checkpoint inhibitors (ICI) and olaparib. The data suggest that gene fusion testing is an important addition to the genomic profiling for therapy selection in solid tumors. Citation Format: Miyoung Shin, Paris Petersen, Steven Lau-Rivera, Segun Jung, Sally Agersborg, Jacyln Hechtman, Fernando Lopez-Diaz, Vincent Funari. Actionable fusions detected by RNA-seq co-occur with PD-L1 expression and driver mutations in solid tumors patients [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 1254.

Abstract 2764: Safety and efficacy of CAR T and TCR therapies in solid tumors: A systematic review and meta-analysis, including a comparison with five phase II trials in hematologic malignancies used for the first FDA approvals of these agents

Abstract Background: Adoptive cellular therapies (ACT) have met with significant success in hematologic malignancies. The number of ACT trials in solid tumors has been increasing, however data are limited on the overall safety and efficacy of ACT in this population. We aimed to conduct a systematic review and meta-analysis looking specifically at TCR and CAR-T in solid tumors and rates of cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) relative to the current approved agents in hematologic malignancies. Methods: We conducted a systematic review of published studies/abstracts from January 2015 up to and including July 2021, selecting those that included ≥ 5 patients. Endpoints included rates and differences in CRS and ICANS. Response rates were evaluated. Statistical tests in the pooled data included χ2 and multivariable logistic regression. Results: This meta-analysis included a total of 1,250 patients [566 patients with solid tumors who had received ACT in phase I, II or I/II clinical trials; a comparison was made with five phase 2 clinical trials in hematologic malignancies used for the first FDA approvals of these agents (JULIET, ZUMA-1, ZUMA-2, TRANSCEND and KARMMA), encompassing 684 heme malignant patients]. Rates of CRS of any grade, CRS ≥ grade 3, ICANS of any grade, and ICANS ≥ grade 3 in solid tumors were 33.0%, 3.8%, 2.3% and 2.1%, respectively. Fever, hypotension, and hypoxia were experienced by 37.4%, 18.5%, and 8.5% of solid tumor patients, respectively. CRS, ICANS, fever and hypoxia in solid tumor patients did not differ by whether the ACT received was TCR or CAR T. Objective response (ORR = CR + PR) in patients with solid tumors was higher with TCR compared with CAR-T [odds ratio (OR) = 3.1, P=0.001] and with NY-ESO-1 than other targets (OR=2.0, P=0.031). Compared to hematologic patients treated with ACT, solid tumor patients treated with adoptive cellular therapies were significantly less likely to experience CRS of any grade (62.4% vs. 33.0%, P&amp;lt;0.001), CRS ≥ grade 3 (7.3% vs. 3.8%, P=0.043), fever (50.3% vs. 37.4%, P=0.001), hypoxia (34.1% vs. 8.5%, P&amp;lt;0.001) or hypotension (29.5% vs. 18.5%, P=0.006), ICANS of any grade (29.7% vs. 2.3%, P&amp;lt;0.001), or ICANS ≥ grade 3 (13.9% vs. 2.1%, P&amp;lt;0.001). However, objective response (20.1% vs. 75.4%, P&amp;lt;0.001) and clinical benefit (CBR = CR + PR + SD) (62.6% vs. 90.9%, P&amp;lt;0.001) rates for ACT were lower in patients with solid tumors than in patients with hematologic malignancies. Conclusions: CRS and ICANs were less prevalent in solid tumor cellular therapies. Based on this analysis, T-cell therapies in solid tumors are relatively safe, but further studies with larger populations and longer follow up are needed to address the role of these treatments in cancer. Citation Format: Mirella Nardo, Ji Y. Son, Goldy G. George, David S. Hong. Safety and efficacy of CAR T and TCR therapies in solid tumors: A systematic review and meta-analysis, including a comparison with five phase II trials in hematologic malignancies used for the first FDA approvals of these agents [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 2764.

Abstract 1774: Circulating IL-6, an early biomarker in HR-positive, HER2-negative metastatic breast cancer patients progressing on CDK4/6 inhibitors

Abstract Background: CDK4/6 inhibitors (CDK4/6i, e.g. palbociclib) in combination with endocrine therapy (ET) have proven successful in delaying progression in hormone receptor-positive (HR+), HER2-negative (HER2-) metastatic breast cancer patients; however, 60-70% will progress within 1-2 years of treatment. Thus, understanding the mechanisms of resistance and identifying novel treatment strategies are needed to improve survival. Our previously published data shows that palbociclib resistance is marked by significant upregulation of the IL6-STAT3 signaling pathway in HR+, HER2- breast cancer cells and treatment with TTI-101, a small-molecule STAT3 inhibitor completing Phase I studies in solid tumor patients, significantly decreased cell viability. Additionally, matched biopsies from advanced HR+, HER2- breast cancer patients who progressed on palbociclib plus ET had an upregulation in activated STAT3 (STAT3 phosphorylated on Y705, pY-STAT3) as compared to their pre-treatment biopsy. Collectively, these data suggest that targeting the IL-6/STAT3 pathway is a viable therapeutic strategy. Our ongoing studies are directed at delineating the role of IL-6 and pY-STAT3 as predictive biomarkers to provide preclinical rationale for targeting STAT3 in patients who develop CDK4/6i resistance. Methods: We downregulated IL-6 using shRNA in MCF7 and T47D palbociclib sensitive and resistant cells. We tested the efficacy of TTI-101 in patient-derived xenograft (PDX) models from patients with metastatic HR+, HER2- breast cancer who progressed on treatment with palbociclib + ET. We also collected plasma samples from patients with metastatic HR+, HER2- breast cancer just prior to CDK4/6i treatment and at disease progression. Results: Downregulation of IL-6 in palbociclib resistant cells reduced pY-STAT3 levels and re-sensitized cells to ET and palbociclib. Induction of IL-6 is an early event by treating sensitive cells with 1uM palbociclib for 21 days during which the cells adapted and eventually developed resistance. Suppression of IL-6 in sensitive cells revealed that it is needed for cells to overcome growth inhibition by palbociclib. Targeting STAT3 in palbociclib resistant PDX mouse models using TTI-101 (50mg/kg twice daily by oral gavage), resulted in significantly reduced tumor volume; a synergistic effect was observed when TTI-101 was combined with palbociclib. Lastly, IL-6 levels in plasma samples from metastatic HR+, HER2- breast cancer patients were significantly higher at disease progression compared to pre-treatment with CDK4/6i. Conclusion: Collectively, these studies suggest that IL-6 signaling through STAT3 is a key mediator of resistance to palbociclib. We provide the rationale for using circulating IL-6 as a biomarker of disease progression and tumor pY-STAT3 as a selection criterion for treatment with TTI-101 to prevent or reverse resistance. Citation Format: Nicole M. Kettner, Tuyen N. Bui, Juliana Navarro-Yepes, T Kris Eckols, Akshara S. Raghavendra, David J. Tweardy, Kelly K. Hunt, Debu Tripathy, Khandan Keyomarsi. Circulating IL-6, an early biomarker in HR-positive, HER2-negative metastatic breast cancer patients progressing on CDK4/6 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 1774.

Abstract 601: IO-108, A fully human therapeutic antibody blocking the myeloid checkpoint LILRB2/ILT4, promotes innate and adaptive anti-cancer immunity in preclinical studies

Abstract Tumor-associated myeloid cells inhibit anti-cancer immune responses systemically and in the tumor microenvironment (TME), thereby limiting the efficacy of T cell checkpoint inhibitors. However, the plasticity of myeloid cells may enable therapeutic intervention. The immune inhibitory receptor LILRB2 (also known as ILT4) is expressed primarily by myeloid cells (monocytes, macrophages, dendritic cells and neutrophils), and has emerged as a key myeloid checkpoint contributing to the tolerogenic activity of myeloid cells associated with cancer. LILRB2 has several ligands (classical and non-classical MHC-I, ANGPTL2/5, SEMA4A and CD1) and most of these are known to contribute to immune suppression in the TME. Using computational biology approaches, we found that high LILRB2 expression is associated with macrophage infiltration in many solid tumor types from TCGA. Therefore, LILRB2 is a compelling target to overcome immune suppressive activity of cancer-associated myeloid cells. IO-108 is a fully human IgG4 therapeutic antibody that binds to LILRB2 with high affinity and specificity. IO-108 binds to all myeloid cells in the solid TME and periphery. In vitro studies support that blockade of LILRB2 interaction with its various ligands is the anticipated mechanism of action of IO-108. The LILRB2 antagonist activity of IO-108 produces the desired pro-inflammatory (re)polarization of myeloid cells. IO-108 treatment results in increased pro-inflammatory responses and an enhanced antigen-presenting cell (APC) phenotype to multiple stimuli (e.g., T cell activators, STING and TLR agonists) in ex vivo assays. In addition, IO-108 precludes the anti-inflammatory myeloid cell phenotype resulting from “tumor conditioning” and promotes the differentiation of monocytes and immature dendritic cells (DC) into pro-inflammatory DC. IO-108 enhances the effect of PD-1 blocking antibodies in allogeneic mixed leukocyte reactions of CD4+ T cells and macrophages. Moreover, IO-108 polarized primary myeloid cells isolated from solid tumor patient blood and ovarian cancer-associated ascites towards a proinflammatory phenotype and attenuated their suppressive effect on autologous T cell proliferation and production of proinflammatory cytokines. In vivo, IO-108 inhibits tumor growth in mouse models, which is associated with immune cell activation. Importantly, IO-108 presents a favorable pharmacokinetic and safety profile in preclinical models. Collectively, the preclinical characterization of IO-108 enabled a comprehensive clinical biomarker plan and lends rationale to the clinical study (NCT05054348) of IO-108 as a novel immunotherapy for multiple solid tumor types, including those relapsed/refractory to standard of care therapies. Citation Format: Jing-Tyan Ma, Xiaoye Liu, Ryan Stafford, Krista McCutcheon, Heyu Chen, Tao Huang, Hongyu Tian, Kyu Hong, ChengCheng Zhang, Luke Chung, An Song, Paul Woodard, XCharlene Liao, MariaJose Costa. IO-108, A fully human therapeutic antibody blocking the myeloid checkpoint LILRB2/ILT4, promotes innate and adaptive anti-cancer immunity in preclinical studies [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 601.

Abstract 1924: PULS-AI: A multimodal artificial intelligence model to predict survival of solid tumor patients treated with antiangiogenics

Abstract The need for developing new biomarkers is increasing with the emergence of many targeted therapies. In this study, we used artificial intelligence (AI) to develop a multimodal model (PULS-AI) predicting the survival of solid tumor patients treated with antiangiogenic treatments. Our retrospective, multicentric study included 616 patients with 7 different cancer types: renal cell carcinoma, colorectal carcinoma, hepatocellular carcinoma, gastrointestinal carcinoma, melanoma, breast cancer, and sarcoma. A set of 196 patients was left out of the study for validation. Clinical data including patient, treatment, and cancer metadata were collected at baseline for all patients, as well as computed tomography (CT) and ultrasound (US) images. Radiologists annotated all metastases on the CT images and the visible tumor lesion on the US images. AI models were used to extract relevant features from the regions of interest on CT and US images. In addition, handcrafted features related to the tumor burden were extracted from the annotations of all lesions on CT such as the number of lesions and the tumor burden volume per organ (lungs, liver, skull, bone, other). Finally, a Cox regression model was fitted to the set of imaging features and clinical features. The annotation process led to 1147 annotated US images with lesions delineation and 4564 reviewed CTs, of which 989 were selected and fully annotated with a total of 9516 annotated lesions.The developed model reaches an average concordance index of 0.71 (0.67-0.75, 95% CI). Using a risk threshold of 50%, PULS-AI model is able to significantly isolate (log-rank test P-value &amp;lt; 0.001) high-risk patients from low-risk patients (respective median OS of 12 and 32 months) with a hazard ratio of 3.52 (2.35-5.28, 95% CI). The results of this study show that AI algorithms are able to extract relevant information from radiology images and to aggregate data from multiple modalities to build powerful prognostic tools. Such tools may provide assistance to oncology clinicians in therapeutic decision-making. Citation Format: Kathryn Schutte, Fabien Brulport, Sana Harguem-Zayani, Jean-Baptiste Schiratti, Ridouane Ghermi, Paul Jehanno, Alexandre Jaeger, Talal Alamri, Raphael Naccache, Leila Haddag-Miliani, Teresa Orsi, Jean-Philippe Lamarque, Isaline Hoferer, Littisha Lawrance, Baya Benatsou, Imad Bousaid, Mickael Azoulay, Antoine Verdon, François Bidault, Corinne Balleyguier, Victor Aubert, Etienne Bendjebbar, Charles Maussion, Nicolas Loiseau, Benoit Schmauch, Meriem Sefta, Gilles Wainrib, Thomas Clozel, Samy Ammari, Nathalie Lassau. PULS-AI: A multimodal artificial intelligence model to predict survival of solid tumor patients treated with antiangiogenics [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 1924.

Abstract 1238: Comprehensive ctDNA monitoring provides early signal of clinical benefit with a novel personalized neoantigen directed immunotherapy for late-stage cancer patients

Abstract Neoantigen directed immunotherapy holds promise to increase the likelihood of patients with solid tumor devoid of immune infiltration benefiting from immune checkpoint immunotherapy (CPI). A heterologous prime-boost vaccination approach consisting of Chimpanzee Adenovirus (ChAd) prime and multiple self-amplifying mRNA (SAM) boosts, delivering 20 neoantigens, has been evaluated in a Phase 1/2 clinical trial in late-stage solid tumor patients in combination with nivolumab and ipilimumab (NCT03639714). Neoantigen dynamics, tumor burden and genomic correlates of response were studied over time in 20 patients (8 GEA, 2 NSCLC, 10 MSS-CRC). Exome sequences from archival (sample used for neoantigen selection), baseline (start of immunization) and on-treatment biopsies were analyzed for 20, 16 and 10 patients respectively. Paired pre- and post-vaccine tumor transcriptomes were analyzed for 6 patients. Personalized capture baits were designed for all non-synonymous mutations detected in archival biopsies (mean 146; range: 67-402) for ctDNA monitoring. Longitudinal ctDNA samples were collected monthly on treatment (mean 7; range: 1-18). ctDNA duplex UMI libraries were captured and sequenced to a target mean raw depth &amp;gt;80,000x and reduced to 3x per strand consensus duplex reads. The majority of vaccine neoantigens were detected in ctDNA (87%; range 45%-100%) and mean neoantigen variant allele frequency (VAF) strongly correlated with all monitored mutations VAF (R2 = 0.90, p &amp;lt; 0.0001) through treatment. The percentage of vaccine neoantigens detected was higher than that for all monitored mutations in the same samples with a median of 80% (21%-98%) in ctDNA and 70% (44%-100%) in biopsies. Five of 9 MSS-CRC patients with measurable baseline ctDNA achieved molecular responses (MR, &amp;gt;50% reduction in ctDNA from baseline) that correlated with OS and PFS, and in some patients, was accompanied by radiologic tumor shrinkage. One MSS-CRC patient with MR for &amp;gt;7 months acquired novel biallelic loss-of-function mutations in TAP1 following 1 year of study treatment. Differential gene expression analysis from paired pre- and post-vaccine biopsies (including 2 MSS-CRC pairs with MR) demonstrated significant upregulation in gene signatures associated with immune-inflamed tumor microenvironments including interferon alpha and gamma responses. We demonstrate that tumor-informed neoantigen selection and vaccine manufacturing while patients receive chemotherapy is feasible, since the majority of neoantigens are retained in the tumor post-chemotherapy. Further, neoantigen-directed immunotherapy appears to drive clinical benefit in patients with advanced MSS-CRC tumors, where CPI alone has provided minimal benefit. Comprehensive ctDNA longitudinal monitoring enables real time assessment of clinical response and acquired resistance. Citation Format: Desiree Schenck, Rita Zhou, Alexis Mantilla, Oliver Spiro, Taylor Patch, Adrienne Johnson, Daniel Navarro Gomez, Brian S. Henick, Chih-Yi Liao, Sameek Roychowdhury, Steve Maron, Benjamin Solomon, Alexander I. Spira, Daniel V. Catenacci, Andrew R. Fergusson, Raphael F. Rousseau, Karin Jooss, Matthew J. Davis. Comprehensive ctDNA monitoring provides early signal of clinical benefit with a novel personalized neoantigen directed immunotherapy for late-stage cancer patients [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 1238.

Abstract CT142: GD2.Ox40.CD28.z CAR T cell trial in neuroblastoma and osteosarcoma

Abstract Background: Chimeric antigen receptor T cells (CARTs) hold promising therapeutic potential for refractory tumors. GD2 is a tumor antigen expressed on neuroblastoma and osteosarcoma. In initial studies, T cells expressing 1st generation GD2-CARTs were shown to be safe and mediated modest antitumor activity in some patients with refractory neuroblastoma. Methods: We developed a 3rd generation GD2-CART (GD2-CAR.OX40.28.z.ICD9) and conducted a phase I trial (NCT02107963) to determine the feasibility of producing and safety of administering escalating doses of GD2-CARTs in children and young adults with GD2+ solid tumors, including neuroblastoma and osteosarcoma, following cyclophosphamide-based lymphodepletion. Patient samples were evaluated for cytokine profile kinetics, immune phenotype analysis with mass cytometry (CyTOF), transcriptomic evaluation with RNA-sequencing (RNA-seq), epigenetic determination with Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq), and functional studies with flow cytometry. Results: 15 patients aged 8-28 years were enrolled on four dose levels, of which 13 patients were infused. No dose-limiting toxicities were observed and administration of up to 1x107 GD2-CART/kg was feasible and safe for children and young adults with neuroblastoma and osteosarcoma. 15.4% (2/13) of patients experienced grade-1 cytokine release syndrome (CRS) and no neurological toxicity was observed. We measured the expansion and persistence of adoptively transferred GD2-CARTs in the peripheral blood. GD2-CARTs expanded in all patients receiving treatment, half of whom had expansion similar to that seen in clinically active CD19 and CD22 CARTs, but the GD2-CARTs had limited persistence. At Day 28 following GD2-CART infusion, 23.1% (3/13) of evaluable patients had progressive disease and 76.9% (10/13) had stable disease (SD). 3/10 SD patients remained stable at 60 days post-GD2-CART, but all patients eventually progressed. Since a major barrier to CART efficacy is inadequate CART expansion, we comprehensively evaluated for phenotypic, transcriptomic, and epigenetic immune profiles of patient apheresis, CART product, and post-treatment peripheral blood samples to identify determinants of CART expansion. GD2-CART expansion is significantly correlated with several T cell markers, and a larger baseline naïve and central memory T cell pool. Unique myeloid populations are associated with CART expansion. ATACseq identifies epigenetic differences in pre-treatment apheresis that may predict good CAR expansion in patients. Conclusions: GD2-CART therapy following cyclophosphamide conditioning was well tolerated at all four dose levels in pediatric and young adult patients with neuroblastoma and osteosarcoma. Subsequent multi-dimensional analyses suggest key mechanisms underlying CART biology and function and highlight the potential of defining and applying molecular signatures in apheresis and CART product as biomarkers and prognostic indicators of CART expansion, with promise for advancing immunotherapies for solid tumor patients in the future. Citation Format: Sneha Ramakrishna, Sabina Kaczanowska, Tara Murty, Cristina F. Contreras, Melinda Merchant, John Glod, Norma Gutierrez, Ahmad Alimadadi, David Stroncek, Steven Highfill, Caroline Duault, Priyanka B. Subrahmanyam, Tyson Holmes, Warren Reynolds, Reema Baskar, Antoine Barge, Hayley Lyon, Radim Moravec, Srinika Ranasinghe, Joyce Yu, Roshni Biswas, Samuel Pollack, Stephen Van Nostrand, James Lindsay, Mina Pichavant, Bita Sahaf, Sean C. Bendall, Andrew J. Gentles, Holden Maecker, Catherine C. Hedrick, Crystal Mackall, Rosandra Kaplan. GD2.Ox40.CD28.z CAR T cell trial in neuroblastoma and osteosarcoma [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 CT142.

Abstract 5844: A comprehensive pan-cancer analysis of MDM2/MDM4 gene amplification in Chinese solid cancer patients

Abstract Background: Many patients with cancer have benefited from immunotherapy represented by the blockade of immune checkpoint PD-1/PD-L1. However, about 4-29% of cancer patients treated with immunotherapy developed hyper-progressive diseases(HPD) with accelerated tumor growth. MDM2/MDM4 amplification was suggested as genomic correlates for elevated risk hyperprogression. The objective of this study was to evaluate the frequency of amplification of MDM2/MDM4 in the Chinese solid tumor patients and to provide advice on the selecting of optimal treatment decisions. Methods: Tumor or blood samples were collected from 2,937 patients with solid cancer, genetic mutations and amplification was detected using the next-generation sequencing(NGS). Results: A total 112 patients with MDM2/MDM4 amplification were identified, including lung cancer 50.9% (57/112), colorectal cancer 14.3% (16/112), gastric cancer 8.0% (9/112), liver cancer 5.4% (6/112), Biliary tract cancers 4.5% (5/112), breast cancer 2.7% (3/112), and pancreatic ductal adenocarcinoma 2.7% (3/112). TMB value was identified, The median TMB for lung cancer was 9.13 mutations/Mb, median TMB for colorectal cancer was 13.8 mutations/Mb, a high TMB value in gastric cancer (median TMB=20.22 mutations/Mb), and median TMB for liver cancer was 17.9 mutations/Mb. Meanwhile, MSI was detected in 8% of patients with MDM2/MDM4 amplification, of which 3 in lung cancer, 5 in colorectal cancer, and 1in gastric cancer. Conclusions: Although amplification of MDM2/MDM4 was observed only in a small proportion of patients, it demonstrated an association with high TMB in several common cancer types. In addition, our data indicate that amplification of MDM2/MDM4 in different types of cancer is heterogeneous and that an appropriate immunotherapy strategy for patients with solid cancer should be noted. Citation Format: xiangyu sun, Zhichao Fu. A comprehensive pan-cancer analysis of MDM2/MDM4 gene amplification in Chinese solid cancer patients [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 5844.

PPM1D in Solid and Hematologic Malignancies: Friend and Foe?

In the face of constant genomic insults, the DNA damage response (DDR) is initiated to preserve genome integrity; its disruption is a classic hallmark of cancer. Protein phosphatase Mg2+/Mn2+-dependent 1D (PPM1D) is a central negative regulator of the DDR that is mutated or amplified in many solid cancers. PPM1D overexpression is associated with increased proliferative and metastatic behavior in multiple solid tumor types and patients with PPM1D-mutated malignancies have poorer prognoses. Recent findings have sparked an interest in the role of PPM1D in hematologic malignancies. Acquired somatic mutations may provide hematopoietic stem cells with a competitive advantage, leading to a substantial proportion of mutant progeny in the peripheral blood, an age-associated phenomenon termed "clonal hematopoiesis" (CH). Recent large-scale genomic studies have identified PPM1D to be among the most frequently mutated genes found in individuals with CH. While PPM1D mutations are particularly enriched in patients with therapy-related myeloid neoplasms, their role in driving leukemic transformation remains uncertain. Here, we examine the mechanisms through which PPM1D overexpression or mutation may drive malignancy by suppression of DNA repair, cell-cycle arrest, and apoptosis. We also discuss the divergent roles of PPM1D in the oncogenesis of solid versus hematologic cancers with a view to clinical implications and new therapeutic avenues.

Analysis of MET genetic aberrations in Chinese solid tumor patients.

e15031 Background: Mesenchymal-epithelial transition (MET) gene encodes a member of the receptor tyrosine kinase family of proteins and is a proto-oncogene. MET gene takes part in various signaling pathways such as PI3K/AKT, Ras/MAPK, and JAK/STAT which are associated with tumor development and progression. This study aimed to reveal MET aberrated characteristics in Chinese solid tumor patients and explore the occurrence of MET oncogenic variants in solid tumors. Methods: This study retrospectively analyzed MET alterations in 1856 solid tumor patients in China from laboratory of Simcere Diagnosis (Nanjing, China) during 2019-2021. Next-generation sequencing (NGS) was performed to detect gene mutations in tumor or blood samples of the patients. Results: We screened out 1856 patients with MET gene alterations from 45717 patients with solid tumor. The high frequency of MET gene alterations was lung cancer (72.8%), colorectal cancer (10.7%), gastric cancer (4.9%), and liver cancer (2.7%). MET gene variants included missense (61.7%), copy number variation (copy number≥5) (15.4%), truncation (7.9%), splicing site variant (7.2%),fusion (1.7%),and in-frame insertion/deletions (0.9%). Among them, MET amplification was detected in 285 patients, and 93 patients carried high-level MET amplification (copy number≥10). High-level MET amplification was most frequently detected in lung cancer (41, 44.1%), gastric cancer (17, 18.3%) and liver cancer (14, 15.1%). MET exon 14 skipping was detected in 287 solid tumor patients (15.5%), of which 277 patients were lung cancer, the remaining were liver cancer, colorectal cancer, gastric cancer, osteosarcoma, melanoma, bladder cancer and glioma. We analyzed MET exon 14 alterations and classified them into 4 subgroups according to the alteration types and locations. The most common alterations were at splice donor sites (215, 74.91%), followed by the splice acceptor sites (35, 12.20%), Y1003 site (5, 1.74%), and other types (32, 11.15%). Other types included insertion or deletion in MET intron 13 or 14, for example, c.3028+21_3028+65del, c.2888-20_2888-11del, c.2888-27_2888-9del, etc. 7 lung cancer patients carried MET exon 14 skipping and MET amplification simultaneously. 36 patients carried MET gene fusions, and mostly were lung cancer (22, 61.1%). The breakpoints often located in exon 2 of MET (4, 11.1%). Conclusions: We discovered MET alterations in multiple types of solid tumors in this study. We found classic MET exon 14 skipping and MET high-level amplification in solid tumors except for lung cancer. It suggested that the activation of MET-driven signaling pathway was ubiquitous in pan-cancer, which might benefit from MET inhibitors.

Solid tumor patients with G12V and G13D KRAS aberrations have poor survival following ICI treatment.

e14567 Background: KRAS gene has been well known as a dominant oncogene which activates downstream signaling for tumor growth. The new therapeutic drug targeting specific KRAS mutation (G12C) has been developed and several clinical trials are testing KRAS inhibitors in solid tumor patients. In line with this, we have surveyed the prevalence and clinical features of solid tumor patients with KRAS aberration in 1,744 oncology patients who received next generation sequencing (NGS) in a clinical practice. Methods: Between 2019 amd 2021, 1,744 oncology patients have received NGS (TruSight Oncology 500 panel; 523 genes) at Samsung Medical Center. . Matched clinical and histology outcomes including programmed cell death-ligand 1 (PD-L1) 22C3 expression (n = 798) and immunotherapy treatment outcomes (n = 409) were analyzed based on genomic alteration. Results: Among the thirty-three different cancer types, the most common cancer types were colorectal cancer (CRC) (N = 547, 31.4%), followed by gastric cancer (GC) (N = 381, 21.8%), and sarcoma (N = 155, 8.9%). Of 1,744 patients, any KRAS aberrations (CNVs, SNVs) were detected in 487 (27.9%) patients (CNV: Copy Number Variation, SNV: Small Nucleotide Variation). Of the 51 (2.9%) patients with KRAS amplification, 18 (35.3%) patients were CRC followed by GC (N = 9, 17.6%). Each 4 cases were found in Cholangiocarcinoma (CCC), sarcoma, melanoma and pancreatic cancer (8%, respectively). Copy number of KRAS were distributed from 4 to over 50, while bladder cancer had the highest median value of copy number(median: 148), followed by CCC(median: 16.2). Of 454 SNV cases, the most frequent cancer type was CRC (N = 259, 57.0%) and pancreatic cancer (N = 89, 19.6%). Except 4 cases, all SNV mutation type was missense mutation. G12D amino acid change was the most frequent(N = 182), followed by G12V(N = 100), G13D(N = 48), and G12C(N = 20). However, only the patients with G12V and G13D mutation had significantly shorter OS and PFS in ICI treatment. Conclusions: Based on our comprehensive survey of next generation sequencing focused on KRAS aberration, we identified approximately 2.9% KRAS amplification and 26.0% KRAS SNV in patients with metastatic solid tumors. It was demonstrated that G12V, G13D were important mutations which were related to the response to ICI treatment in real-world data.[Table: see text]

Utilization and financial coverage of blood based next generation sequencing for solid tumors: A study from Augusta-Georgia.

e18592 Background: There is an increasing trend of integrating molecular profiling to devise treatment plans for solid malignancies. Tumor testing for specific clonal abnormalities is being supplanted by broad multigene panel testing from blood specimen, aptly labelled liquid biopsy. Due to varying utility across malignancies in differing contexts and issues of access and affordability, their adoption is not uniform. We attempt to characterize the pattern of utilization across tumor types and financial coverage schemes in an academic cancer institute in southeastern United States. Methods: All blood based next generation testing samples ordered by providers affiliated to Georgia Cancer center between January 1, 2017, and December 31, 2021, via Guardant platform for all solid tumors was included in this analysis. Guardant360 and Guardant360CDx assays tested for upto 83 cancer related genes in circulating cell-free DNA in patients of solid tumors. Primary cancer diagnosis, insurance information and turnaround time was obtained from Guardant. Results: 296 tests were sent from January 1, 2017, through December 31, 2021. The total number of tests performed in year 2017, 2018, 2019, 2020 and 2021 were 1, 3, 28, 99 and 165 respectively. Billing information was available for all cases, clinical information was available for 262 cases. Average turnaround time was 8.5 days overall. 221 Guardant360 was sent with average turnaround time of 10 days. 41 Guardant360CDx was sent with average turnaround time of 7 days. The frequency of malignancies in the order of dimensioning frequency were lung 50.7%, prostate 12.1%, breast 6.9%, kidney 3.8% and colorectal 3.4%. The remaining 22.1% comprised of pancreatic, melanoma, glioblastoma, and bladder malignancies. The test was covered by private insurance/employee health plan in 53.4% (158/296), Medicare in 31.1% (92/296) and Medicaid in 10.5% (31/296) (table). 4.7% (14/296) qualified for free of charge testing (financial indigent) and 0.3% (1/296) were self-pay (table). Conclusions: There is an increase in the utilization of liquid biopsy to perform comprehensive genomic analysis. The distribution of tumor types is not entirely explained by the incidence and availability of targeted therapies for each type. Private insurers accounted for most of the tests being done. The disparities in utilization of this platform across different tumor types and payment models deserve further exploration. T[Table: see text]

Safety, tolerability, and preliminary efficacy of nadunolimab, a first-in-class monoclonal antibody against IL1RAP, in combination with pembrolizumab in subjects with solid tumors.

2527 Background: Interleukin-1 Receptor Accessory Protein (IL1RAP) is expressed on cancer and stromal cells of many solid tumors. IL1RAP interacts with IL-1R1, modulating downstream factors (e.g. IL-6, IL-8) and CRP level. Nadunolimab (CAN04), a fully humanized ADCC-enhanced IgG1 antibody, targets IL1RAP and blocks IL-1α and IL-1β. IL-1 promotes an immune suppressive microenvironment, e.g. by recruitment of MDSC which may induce checkpoint inhibitor resistance. Here, initial data are reported from the phase Ib clinical trial CIRIFOUR, evaluating nadunolimab combined with pembrolizumab in solid tumor patients (pts) progressed on prior anti-PD-(L)1 therapy. Methods: Primary objective was evaluating safety and tolerability of nadunolimab combined with pembrolizumab in pts with metastatic NSCLC (n = 5), head and neck squamous cancer (n = 9) or malignant melanoma (n = 1), progressed on prior anti-PD-(L)1 therapy of ≥12 weeks (wks). Dosing started with a priming dose of 0.5 mg/kg nadunolimab on Day -7, followed by 5 mg/kg nadunolimab weekly combined with standard pembrolizumab dosing. A safety lead-in phase 3+3 design was employed. Secondary objectives included preliminary efficacy and responses based on RECIST/iRECIST. Serum biomarkers and tumor biopsies were also analyzed. Results: Fifteen pts received at least one dose of 5 mg/kg nadunolimab with pembrolizumab: median age 63 years (50-79), 27% female, 100% Stage IV, 93% and 7% received prior pembrolizumab or nivolumab respectively, 7% ECOG 0, 93% ECOG 1. The pts had received a mean of 2.3 (range 1-6) previous lines of treatment. SAE were reported in 47% with one considered treatment-related (febrile neutropenia). Two treatment-related grade 3 AE were reported: one febrile neutropenia (DLT), one pneumonitis. No pts discontinued treatment due to treatment-related AE. One patient (7%) had unconfirmed PR as best response, 8 (53%) showed SD, and 6 (40%) iUPD. At time of analysis, 5 pts (33%) received ongoing treatment. Of these, two pts had received therapy for over 31 wks, another two for over 49 wks. Decreased IL-6 was observed after four wks and persisted during treatment. Reduced neutrophil-lymphocyte ratio (NLR) was observed throughout the study, appeared after first treatment, and was driven by a moderate reduction in circulating neutrophils. Decreased IL-6 and NLR were most pronounced in pts with longest disease control duration. IL1RAP expression on cancer and stromal cells was confirmed in tumor biopsies. Conclusions: The nadunolimab and pembrolizumab combination was considered safe and tolerable with preliminary evidence of prolonged disease control. The favorable safety provides basis for evaluation of further therapy with this combination. Next, nadunolimab and pembrolizumab will be assessed with carboplatin/pemetrexed in non-squamous NSCLC. Clinical trial information: NCT04452214.

A pan-cancer analysis of P21 activated kinase (PAK) family genes as potential biomarkers for immune checkpoint therapy.

2614 Background: PAK (P21 activated kinase) is a family of serine threonine kinases, that consist of 6 members, PAKs 1-6, which are positioned at an intersection of multiple signaling pathways implicated in oncogenesis. Recent studies have found relationship between mutations of some PAK family genes and antitumor immunity. Here, we explored the associations of PAK family gene mutations with ICI response based on multidimensional data from multiple solid tumors. Methods: An immunotherapy cohort (Broad/Dana-Farber, Nat Genet 2018, N = 249) within 7 types of tumors as discovery stage, was used to explore the association of PAK family genes mutations with tumor mutation burden (TMB) and efficacy of immunotherapy. To further validate the predictive value of PAK family genes, we employed another ICI-treated cohort, MSKCC cohort (Samstein, Nat Genet 2019, n = 1661), as a validation cohort. TMB was calculated as the total count of nonsynonymous mutations in coding sequence. Results: In discovery cohort, compared to that in the PAK-Wt group, longer OS was observed in the PAK-Mut group (mOS: 31.3 vs 15.4 months, HR = 0.59, 95% CI: 0.36 to 0.96, P= 0.03). In addition, PAK-Mut group had higher TMB ( P&lt; 0.001). Median TMB in PAK-Mut group and PAK-Wt group is 17.79 (IQR, 7.45-36.55) Mut/Mb and 5.47 (IQR, 2.46-9.09) Mut/Mb, separately. In validation cohort, PAK-Mut patients also achieved significant improved OS compared with PAK-Wt patients (mOS: 44.0 vs 17.0 months, HR = 0.51, 95% CI: 0.37 to 0.69, P&lt; 0.001). Moreover, multivariable analysis of validation cohort demonstrated that PAK-Mut was associated with better OS (HR = 0.71; 95%CI, 0.52-0.98; P = 0.036), after adjusting for Age, Gender, Metastasis, Treatment, TMB, Cancer type. Compared with PAK-Wt group (median [IQR]: 5.90 [2.95-10.04]), PAK-Mut group (median [IQR]: 21.19 [11.81-43.29]) also had higher TMB ( P&lt; 0.001). Conclusions: The results indicated that PAK family genes mutation was associated with a higher TMB both discovery and validation cohort. Analysis of discover and validation immunotherapy cohort data showed PAK family was associated with better OS. These findings indicate that these genes mutation may serve as a potential predictive biomarker for ICI in solid tumor patients.