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AML-355: The Prognostic Validity of the Acute Myeloid Leukemia Composite Model in Predicting Risks of One-Year Mortality among Patients in Atlantic Canada: A Multi-Centre Experience
Abstracts widely varying target expression. Keywords: single-cell analysis, apoptosis, CAR therapy, BCL-2, T-cell exhaustion, PD-1, AML, acute myeloid leukemia
AML-355 The Prognostic Validity of the Acute Myeloid Leukemia Composite Model in Predicting Risks of One-Year Mortality among Patients in Atlantic Canada: A Multi-Centre Experience
Keywords: leukemia, comorbidities, induction, AML, AMLcomposite model
AML-356 High Dimensional Analytic Approaches Identify Stressor Specific Response Patterns in AML Muharrem Muftuoglu*, Po Yee Mak, Vivian Ruvolo, Yuki Nishida, Peter Ruvolo, Bing Z. Carter, Michael Andreeff
Shannon Murphy 1*, Brian Harnett 2, Kara Matheson 1, David Jones 2, Abdulrahim Basendwah 3, Sudeep Shivakumar 1, Mahmoud Elsawy 1
Section of Molecular Hematology and Therapy, Department of
1
Cellular response to stress is diverse, ranging from adaptation to induction of cell death. We designed a multicolor flow cytometry panel to gain insight into multifaceted stress response and to assess multiple cell death modes. This enabled simultaneous interrogation of multiple cell death modes including necrosis, necroptosis, apoptosis and parthanatos as well as proliferation, autophagy and endoplasmic reticulum (ER) stress. Notably, we utilized high dimensional analytic approaches to better elucidate stress response and cell death modes. We aimed to delineate response patterns and cell death modes associated with targeted agents in the context of AML. Bcl-2 or Mcl-1 inhibition alone did not alter cellular landscape. However, treatment with MDM2 or exportin 1 (XPO1) inhibitors and mitochondrial proteasome (ClpP) activator (ONC201) elicited divergent stress responses and cell death modes. Two-dimensional UMAP plots showed differential induction of autophagy and ER stress following treatment with MDM2, XPO1 inhibitors/ClpP activators. Remarkably, we observed that MDM2i and XPO1i were associated with emergence of quiescent cells, based on high p21 expression, and higher levels of ER stress and autophagy while ClpPa induced DNA damage, and was associated with persistent Ki-67 expression and lower levels of p21. This approach enabled us to dissect single agent specific stress signatures. Remarkably, dual combinations differentially altered and skewed response patterns. Strikingly, combinatorial use of MDM2i and XPO1i almost completely eliminated all AML cells except the ones with the highest levels of ER stress and autophagy. On the other hand, addition of either Bcl-2i or MCL-1i to MDM2i markedly reduced p21, ER stress and autophagy, indicating that these antiapoptotic molecules may play a role in cellular adaptation. We leveraged diffusion map algorithm to map cellular trajectories. This approach enabled us to identify the earliest stage of cell death, characterized by expression of LC3B, H2AX and cleaved PARP while dead cell dye (+) cells marked the latest stage. These findings provide proof of concept for the utility of single cell mapping of cellular stress in delineating stressor-specific response patterns and identifying potential resistance mechanisms. Single cell mapping of cell stress and cell death can inform the development of more effective combinatorial drug regimens. Keywords: flow cytometry, single-cell analysis, cell death, cell stress, cell trajectory, dimension reduction, clustering, AML
Dalhousie University, Halifax, NS, Canada, 2Memorial University
of Newfoundland, St. John’s, NL, Canada, 3King Fahad Armed Forces Hospital, Jeddah, Saudi Arabia
Background: Intensive chemotherapy is the standard of care for remission induction for fit patients with newly diagnosed acute myeloid leukemia (AML). However, assessing the prognostic impact of comorbidities on outcomes when counselling patients remains a challenge. To this end, we sought to validate the prognostic validity of a newly proposed AML composite model (AML-CM) which incorporates both comorbidities and leukemia-specific features to predict overall mortality following administration of intensive induction chemotherapy. Methods: We retrospectively collected data on patients with newly diagnosed AML who received their first induction chemotherapy in Halifax, NS and St. John’s, NL, Canada. The AML-CM scores were calculated as per previously published guidelines (Sorror et al in 2017). Logistic regression models were performed to analyze 8-week and 1-year mortality and competing risk regression with Fine and Grey model to model overall survival after adjusting for known variables. Results: 194 patients treated January 1, 2009 to December 31, 2017 were identified. Fifty-six percent were males. Median age at induction was 54 years with an age range from 18 to 75 years. Thirty-eight percent of patients were 60 years or older. Molecular/cytogenetic risk per ELN classification was as follows: 23% favourable risk, 37% intermediate risk, 34% high risk, and 6% unknown. The most frequently used regimen was the standard 3+7 regimen (daunorubicin and cytarabine). The AML-CM was predictive of 8-week, odds ratio 2.39 (95%CI 1.095.239) AML-CM 7-9 vs AML-CM 1-4, (p-value 0.0284) and 1-year mortality, odds ratio 11.76 (95% CI 2.45-56.51) AML-CM 10+ vs AML-CM 1-4 (p-value 0.0021). Overall survival was inversely proportional to increasing AML-CM scores (p <0.001). Overall survival at 1 year for patients with AML-CM scores 1-4, 5-6, 7-9, 10 was 50%, 30% 18%, and 2%, respectively (p-value 0.002). Conclusions: The AML-CM is a valid prognosticator for early and late mortality in our patient population. Our findings emphasize the significance of comorbidities assessment prior to induction therapy for AML and the potential use in tailoring targeted interventions to mitigate their risks alongside leukemia treatments. Additionally, the AML-CM could be utilized to adjust for the impact of comorbidities in clinical trials investigating newer intensive AML therapies.
Leukemia, MD Anderson Cancer Center
Clinical Lymphoma, Myeloma & Leukemia September 2020
S207
AML-355 The Prognostic Validity of the Acute Myeloid Leukemia Composite Model in Predicting Risks of One-Year Mortality among Patients in Atlantic Canada: A Multi-Centre Experience
Keywords: leukemia, comorbidities, induction, AML, AMLcomposite model
AML-356 High Dimensional Analytic Approaches Identify Stressor Specific Response Patterns in AML Muharrem Muftuoglu*, Po Yee Mak, Vivian Ruvolo, Yuki Nishida, Peter Ruvolo, Bing Z. Carter, Michael Andreeff
Shannon Murphy 1*, Brian Harnett 2, Kara Matheson 1, David Jones 2, Abdulrahim Basendwah 3, Sudeep Shivakumar 1, Mahmoud Elsawy 1
Section of Molecular Hematology and Therapy, Department of
1
Cellular response to stress is diverse, ranging from adaptation to induction of cell death. We designed a multicolor flow cytometry panel to gain insight into multifaceted stress response and to assess multiple cell death modes. This enabled simultaneous interrogation of multiple cell death modes including necrosis, necroptosis, apoptosis and parthanatos as well as proliferation, autophagy and endoplasmic reticulum (ER) stress. Notably, we utilized high dimensional analytic approaches to better elucidate stress response and cell death modes. We aimed to delineate response patterns and cell death modes associated with targeted agents in the context of AML. Bcl-2 or Mcl-1 inhibition alone did not alter cellular landscape. However, treatment with MDM2 or exportin 1 (XPO1) inhibitors and mitochondrial proteasome (ClpP) activator (ONC201) elicited divergent stress responses and cell death modes. Two-dimensional UMAP plots showed differential induction of autophagy and ER stress following treatment with MDM2, XPO1 inhibitors/ClpP activators. Remarkably, we observed that MDM2i and XPO1i were associated with emergence of quiescent cells, based on high p21 expression, and higher levels of ER stress and autophagy while ClpPa induced DNA damage, and was associated with persistent Ki-67 expression and lower levels of p21. This approach enabled us to dissect single agent specific stress signatures. Remarkably, dual combinations differentially altered and skewed response patterns. Strikingly, combinatorial use of MDM2i and XPO1i almost completely eliminated all AML cells except the ones with the highest levels of ER stress and autophagy. On the other hand, addition of either Bcl-2i or MCL-1i to MDM2i markedly reduced p21, ER stress and autophagy, indicating that these antiapoptotic molecules may play a role in cellular adaptation. We leveraged diffusion map algorithm to map cellular trajectories. This approach enabled us to identify the earliest stage of cell death, characterized by expression of LC3B, H2AX and cleaved PARP while dead cell dye (+) cells marked the latest stage. These findings provide proof of concept for the utility of single cell mapping of cellular stress in delineating stressor-specific response patterns and identifying potential resistance mechanisms. Single cell mapping of cell stress and cell death can inform the development of more effective combinatorial drug regimens. Keywords: flow cytometry, single-cell analysis, cell death, cell stress, cell trajectory, dimension reduction, clustering, AML
Dalhousie University, Halifax, NS, Canada, 2Memorial University
of Newfoundland, St. John’s, NL, Canada, 3King Fahad Armed Forces Hospital, Jeddah, Saudi Arabia
Background: Intensive chemotherapy is the standard of care for remission induction for fit patients with newly diagnosed acute myeloid leukemia (AML). However, assessing the prognostic impact of comorbidities on outcomes when counselling patients remains a challenge. To this end, we sought to validate the prognostic validity of a newly proposed AML composite model (AML-CM) which incorporates both comorbidities and leukemia-specific features to predict overall mortality following administration of intensive induction chemotherapy. Methods: We retrospectively collected data on patients with newly diagnosed AML who received their first induction chemotherapy in Halifax, NS and St. John’s, NL, Canada. The AML-CM scores were calculated as per previously published guidelines (Sorror et al in 2017). Logistic regression models were performed to analyze 8-week and 1-year mortality and competing risk regression with Fine and Grey model to model overall survival after adjusting for known variables. Results: 194 patients treated January 1, 2009 to December 31, 2017 were identified. Fifty-six percent were males. Median age at induction was 54 years with an age range from 18 to 75 years. Thirty-eight percent of patients were 60 years or older. Molecular/cytogenetic risk per ELN classification was as follows: 23% favourable risk, 37% intermediate risk, 34% high risk, and 6% unknown. The most frequently used regimen was the standard 3+7 regimen (daunorubicin and cytarabine). The AML-CM was predictive of 8-week, odds ratio 2.39 (95%CI 1.095.239) AML-CM 7-9 vs AML-CM 1-4, (p-value 0.0284) and 1-year mortality, odds ratio 11.76 (95% CI 2.45-56.51) AML-CM 10+ vs AML-CM 1-4 (p-value 0.0021). Overall survival was inversely proportional to increasing AML-CM scores (p <0.001). Overall survival at 1 year for patients with AML-CM scores 1-4, 5-6, 7-9, 10 was 50%, 30% 18%, and 2%, respectively (p-value 0.002). Conclusions: The AML-CM is a valid prognosticator for early and late mortality in our patient population. Our findings emphasize the significance of comorbidities assessment prior to induction therapy for AML and the potential use in tailoring targeted interventions to mitigate their risks alongside leukemia treatments. Additionally, the AML-CM could be utilized to adjust for the impact of comorbidities in clinical trials investigating newer intensive AML therapies.
Leukemia, MD Anderson Cancer Center
Clinical Lymphoma, Myeloma & Leukemia September 2020
S207