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MLFPM Symposium 2022: Pelin Gundogdu

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Pelin Gundogdu presented her work on a neural network project designed to address the "black box" problem inherent in complex machine learning algorithms, particularly within the sensitive healthcare field. While standard deep learning models offer high performance, their lack of interpretability makes it difficult to understand how they arrive at specific conclusions. To solve this, the team integrated biological knowledge directly into the network architecture by incorporating signaling pathways as circuits in the first hidden layer. This design ensures that the model's calculations are grounded in real cellular operations, where each circuit represents a functional unit with receptors or genes that pass signals to subsequent processes, thereby enhancing the explainability of the results without sacrificing predictive power. The project focuses on three core tasks: cell type prediction, cell type annotation, and interpretation using this integrated biological knowledge. The team utilized two datasets derived from the SCITE paper: a balanced PBMC dataset with seven cell types for prediction analysis, and a melanoma dataset used to test both annotation and interpretation capabilities. Through rigorous experiments involving 100 iterations with certified holdout validation, the network demonstrated high accuracy, precision, recall, and F1 scores across all cell types. The results showed that the model could effectively predict cell types and successfully identify unknown or abnormal samples by clustering them separately, proving that the encoding layer retains sufficient information about cell differentiation even when encountering unseen data types. Beyond standard prediction metrics, the study emphasized the importance of biological validation through novelty detection and local outlier factor analysis. By calculating similarity scores based on the activation of nodes in the first hidden layer, the researchers could determine which samples were abnormal or belonged to unknown cell types compared to the training set. The findings indicated that while performance was slightly lower for specific cell types like NK cells due to limited sample sizes, the overall accuracy remained competitive with established algorithms like SCITE. Furthermore, the analysis confirmed that the biological circuits actively participated in the decision-making process, as the most active pathways for specific cells, such as B cells, aligned perfectly with known biological functions like proliferation and protein secretion. The presentation concluded with a discussion on future directions and responses to audience questions regarding model robustness and parameter sensitivity. Gundogdu highlighted ongoing work on cell type trajectory projects using cancer datasets to understand cellular behavior changes during disease progression. When questioned about the network's performance in distinguishing biologically similar cell types, she noted that while the model sometimes struggles with very close clusters, collaboration with biologists to refine definitions could improve results. Additionally, regarding the hard removal of connections not related to specific circuits, she explained that shuffling these connections led to a significant drop in performance, confirming that maintaining biologically meaningful connections is crucial for the network's success, though they did not explore soft regularization approaches as suggested by the audience.
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so peline is one of our esrs at a Spanish node supervised by joakin and is the final speaker of our first day of the symposium that's ready to go yes so we're looking forward to your talk thank you so much Carson uh hi everyone today I'm going to present you an air Network project we developed and oh so yeah before I started I'm applying mention my advisors are Hawking Isabel and Carlos and I start I started September 2019. so as I mentioned earlier before our project is neural network project and more space this project are really a more complex algorithm stand regular machine learning algorithms and we are using them and this complexity enables us to work a really complicated problem and they produce really well performance however this complexity also causes an issue called black box in Black Box concept and the calculation during training are well known however the interpretability of the results is uncomprehensible and this lack of explainability can cause a problem especially if we are working in healthcare field and for this reason we are trying to we are we we are using we are trying to implement a biological knowledge into Network more specifically a signaling Pathways as Hawking mentioned in his earlier speak and a signaling pathway is a is a cellular level operation in an organism and each each pathway can either call either creates a product or an helper signal for the next process and we are specifically using circuits circuits is the smallest element of the pathway and they have um they have a factored or receptorgin information and we are trying to we are using this information to improve the interpret interpretability of the network the network is uses signal signaling a single cell earnings sequencing data and it's uh it's a predict or annotate cell type information and to summarize uh the the project I'm presenting has three main Concepts uh cell type prediction startup annotation and interpretation so a startup prediction is simple supervised approach it's just predicting cell types cell type annotation is focusing on the hidden layer information and the interpretation part is focusing on this integrated knowledge which is the signaling pathways I'm sorry circuits and our design is fit for nail network with Dance Connection and we are using this biological uh knowledge in the first hidden layer and we are implementing the implementation is applied by using um each node in the first hidden layer as the circuit which means if the genes which is the input layer has no connection with the selected circuits done we are removing the weights of course here and for the data sets um we are using two data sets for this project and before giving more details about data set I would like to mention one paper which called sipet cybet and this paper is important uh for us because both of the data sets are shared both of the data sets is obtained from this paper and we are using exactly exact pre-process step as they did because this paper also shared some results according to their algorithm because this cybit algorithm is one of the well-known algorithm for cell type prediction test and they produce really competitive results as I mentioned we are using exact same versions as they they use and the first data set we called is pbmc it has it is a balanced data set and has seven different style types and the second one second one melanoma there are two subsets for this data set there are they called training and testing uh training a testing has this exact same data set as training however there are one additional cell type which training doesn't don't doesn't have and that's and we are using pbrmc dataset for um cell type prediction analysis and the melanoma data set we are using both interpretation and setup annotation tests the task yes uh the purpose of this analysis is just to understand how well the how well our proposed network is because the purpose is just to get the prediction versus Grant true and for these purposes uh we we design an experiment with 100 iteration by using certified holdout validation and each iteration is uses 70 30 of the data set as a testing data set and these are the seven cell types which has the same exact number of sample size and evolution is um for evaluation we have five metrics accuracy balanced accuracy F1 precision and recall scores in addition to these five metrics we are also generating a rich confusion Matrix uh these two figures is the the evolution Matrix result the top figure here is the uh belongs to these five metrics uh actually for balanced and accuracy balanced are actually uh yes all of them seems similar because uh our data set has is balanced data set however we actually generate a calculating the the this paying attention the distribution of each cell each cell type however we are seeing a similar results but this is not an issue because the data set is balanced and both graphs are taking a value between 1 and 0 1 is for best worst for the zero is the worst score and as we can see here they are quite close to one and this shows us that our network is working for Salter prediction in the bottom figure in addition to General overall picture we also pay attention to to Startup details Precision F1 and recalls course is calculated for each cell type detail and they are all close to one however there are a little bit slight difference for these three cell types and so toxic memorizers and regulator however again like overall scores are able above than 0.7 so I'm sorry 0.8 so in overall picture shows that the prediction for each cell types working well and this is the confusion average confusion Matrix and this is a basic run through versus prediction and the diagonal shows how well the predictions are and for these three cell types uh we are slightly low getting lower performance however in biologically they are quite similar and we as we can see here each verse prediction is below is distributed by is distributed by these three cell types uh as I mentioned um cyber paper shared these three results in the bottom these three algorithms are the most well-known and most commonly used algorithms in SATA prediction tests and our results isn't in the top in the top figure which is quite similar to cybet and we are producing really competitive results as cybed and however in addition to these three algorithms we are aiming to interpret our analysis our Network and for this purpose um after we saw that our predictions are really good our assumption was if we are getting good prediction in output layer then the encoding player which is the last hidden layer of the network should also have the information about the information about the startup differentiation and for this reason we the experiment is started with in in two steps the first step is basic visualization just creating clusters and the second one is novelty detection uh which Define each which is we are trying to find the similarities for samples and then try to understand which samples are relatively um abnormal than the rest of the data set and interpretation is again looking the biological layer which is the first hidden layer in the in our design and more specifically and we are looking the activation scores of each node in the first hidden layer for each cell type which means basically we are just getting the most active notes for each cell types and according to this selected cell types selected nodes we are making the analysis and for example for B cells the most active pathway should relate it with side to side communication prolification protein expression and secretion and our results shows that the most active pathways are related with these functions cell functions and this output provides us that during the startup prediction the biological the integrated biological layer is taking an entire account yes and for next one as I mentioned we have two steps for this analysis and both of the analysis is using encodingly encoding layer yes and the experiment is designed by using 70 percent of the training set here and then after model is trained a full training set and the testing set is used for this experiment and the as I mentioned the purpose of this analysis just to provide a visual proof because we want to see can encoding encoding layer can create a clusters for each cell types one important cell type here is negative cells as you can see here this negative cell doesn't exist in training set and we are trying to while we are trying to create a separate Cloud clusters for this cell type while also getting separate classes for rest of the cell types these are the two results as I imagine 70 percent of the training set is using for the model training and this is the full sample set for uh for the final figure as we can see here it's cell type which are the non-style type for the uh for the model can is creating one cluster for each cell type and for the next one testing and as we can see we all we also is we also can be can see that um one cluster for each cell type but also we are seeing that one separate cluster for unknown setup are unknown cell type and this shows us uh the encoding can be used as a is it is a startup differentiation and for this purpose um we are using a similarity score by from local outlier factor analysis and um and the steps are in this analysis start with calculating the encoding information calculated by similarity score by using encoding information this similar score is um shows how for giving sample uh so this this uh this sequential that for giving sample uh have a half similar this sample for the rest of the data set and it gets a 0 to negative infinite zero if zero if we are close to zero zero it means uh this this giving sample has a relationship or more similar that or more similar to rest of the sample set and the next step is we are by using these similar score We call we generate a distribution plot for each cell types and by using these distributions to chords we are calculating a threshold and this threshold will help will is helping us to decide which samples are gonna assigned as an unknown and which samples are going to execute for our Network and for this if the calculated similar to the score for giving sample is above from this threshold then we are executing our Network and end of this end of this network analysis we are getting a SATA prediction and if the label if the value similarity score is below then this threshold then we are saying that the network didn't see doesn't see this label before so that that means like this is unknown uh this is the final results after following all steps and as we can as you can see here we are getting a high accuracy for all the cell types and also we are getting a high accuracy uh for a no setup identification one issue I would like to address is the performance of NK as you know the neural networks work work better with higher sample set for this status cell type unfortunately we have small set a small sample set that's why the performance is relatively lower than we compared rest and this is the comparison between cybet and our Network and as we can see here we are getting a higher performance except NK and also we are getting a similar performance for um unknown cell type annotation uh the results I showed is still ongoing one of our ongoing projects and it's funding by Europe is several European agency and before uh ending my presentation I also would like to imagine our latest project so we are we are currently working a cell type uh trajectory project there are several studies in this field and basically in this project we are trying to find the cell type Behavior according to uh yes for this state for this project we are using cancer data set because we are trying to understand uh the changes in the cell type when it's called it's became the cancer so the idea is if we are try if we if we see this trajectory which is like the pets we can understand how the cell behavior for the patient or the treatment all the or the cancer stage so these are the the full sample set we are using as you can see there are multiple and yes and we are in the initial stage however we are getting um the results we get so far is what the literature says and we are using a variation of the encoder for this design and also we are using um signaling pathway also in this project thank you so much [Applause] foreign [Music] now time for questions Lucas next question thank you very much billing it was a wonderful talk very clear and I also love not only the content but the progress bar at the top yeah it really helps with anxiety um yeah I think everyone here is the same coffee I'm starting to work a bit with our single seller and async data and uh yeah cell type and annotation and prediction seems to be kind of hard in some situations I was wondering if you know how well this algorithms perform in situations at which these cell types are not that different for example with single nuclear rna-seq in which the different cell types like for example different types of neurons [Music] um for I mean in biological point of view I'm not like that expert but I know there are for example in our result like confusion Matrix if the cell types are really close to each other the network is quite underperformance so yes for this purpose I think I mean in my opinion it worked with some biologists to understand or at least finding a route for the cell types might be might be helped for the network questions so I have one question you you mentioned the local outlier Factor yes um and this includes a number of parameters are your results sensitive these parameters or have you explored this or how do you set them to make yeah actually this was one of the topics when we are using this network tool um how we use the default feature because our aim is to see can be separate or candid default I mean if we are even if we are using simple design can we find this separation between cell types by using the simplest model so just the default parameters in some software implementation of LOF okay good further questions really yeah Giovanni thank you I have a quick question on the structure of the first hidden layer where you mentioned that you keep only the connections for the genes that are relevant to the circuits have you considered doing that instead as a soft version so instead of literally removing the the links between the neurons just adding some regularization term in the uh in your training procedures so that the first layer is incentivized to mimic these connections but also allows for a little bit of wiggle room to try to find something extra do you mean what the regulation do you mean like um I mean L1 L2 regulations no you can just add the term sorry to the loss uh to penalize how far the connection goes from these hard um cutoff but then since it's just determined the loss you could still get a little bit of other contributions and try to get a more General result actually we didn't but we so we make this analysis which is like we we Shuffle the connection you know I mean we assume that I mean we keep the same number of connection but we shuffle them and however when we shuffled them the results we are getting is really full performance but we didn't do anything about what you are saying but when we shuffledam we saw that um this shuffling is really important because the discipline it doesn't mean for the network but keeping the is a biologically meaningful is providing performance thank you are there further questions if not then I would like to think pilling and all the speakers of the first day of this Symposium thank you very much thank you